NIDDK Research Materials

Podocin-rtTA (Tg(NPHS2-rtTA2*M2)1Jbk Mouse

Fri, 20 Mar 2026 09:23:00 -0400

Reverse tetracycline-controlled transgenic mouse model (Tet-On) under the control of the specific promoter for the glomerular podocyte protein (Podocin, NPHS2).

Podocytes are post-mitotic epithelial cells that are positioned on the exterior aspect of the glomerular capillary wall and contribute to the selective molecular permeability of glomeruli. Podocyte damage or dysfunction results in loss of the characteristic foot processes that normally interdigitate and form the selective permeability barriers composed of filtration slits bridged by slit diaphragms. Minimal damage causes proteinuria that, in the case of minimal change disease (MCD), can be reversed by steroid treatment. In focal segmental glomerulosclerosis, more severe loss of podocytes ultimately results in glomerulosclerosis.

The transgenic mouse model described uses a tetracycline controlled reporter-operator system from E. coli to induce the transcription of a target gene in podocytes. Transcription is induced only in podocytes and only when tetracycline is administered. The response is dependent on the tetracycline dose. The podocyte-specific inducible transgene system can be used to identify factors that exacerbate or ameliorate podocyte injury, and can be used to express Cre-recombinase.

In the reverse tetracycline-controlled transcriptional activator system (rtTA), transcription in podocytes is activated by a transgene expressing a chimeric protein containing: (i) the promoter-enhancer region of Nphs2 (podocin, a protein specifically expressed in the podocyte), (ii) a Tet repressor domain that only binds to and activates the tetracycline operator sequence (TetO) when it binds tetracycline, and (iii) a VP16 activation domain. The target gene to be induced is linked to the tetracycline operator (TetO).

Publication

Inducible podocyte-specific gene expression in transgenic mice.: Shigehara T, Zaragoza C, Kitiyakara C, Takahashi H, Lu H, Moeller M, Holzman LB, Kopp JB.; J Am Soc Nephrol (2003 Aug) 14(8):1998-2003. Abstract/Full Text

Inducible podocyte-specific gene expression in transgenic mice

Fri, 20 Mar 2026 09:20:59 -0400

The podocyte plays a key role in glomerular function and glomerular disease. To facilitate studies of podocyte function, we have developed a transgenic mouse model with inducible expression in the podocyte. The tetracycline-inducible transgenic system facilitates gene expression with restricted cellular distribution and tight temporal control. Recently, Bujard and colleagues have developed a functionally improved reverse tetracycline-controlled transcriptional activator (rtTA) with substantially lower background in the off state (the absence of tetracycline) and greater inducibility in the on state (the presence of tetracycline). We used the human podocin (NPHS2) gene promoter to control expression of the rtTA cassette and bred these mice with a reporter mouse line that contains the cytomegalovirus minimal promoter and tetO promoter elements together with LacZ, encoding beta-galactosidase. Dual transgenic mice, bearing both podocin-rtTA and tetO-LacZ transgenes, had no detectable expression in kidney or other organs in the absence of tetracycline. Administration of tetracycline in the drinking water was associated with podocyte expression of beta-galactosidase, in a fashion that was time dependent (maximal at 1 wk) and dose-dependent (maximal at 2 mg/ml). Podocyte expression was confirmed in two ways: histochemical staining for beta-galactosidase and double-immunostaining using the podocyte marker WT-1 and beta-galactosidase. This transgenic system should aid future investigations of podocyte function.

Publication

Inducible podocyte-specific gene expression in transgenic mice.: Shigehara T, Zaragoza C, Kitiyakara C, Takahashi H, Lu H, Moeller M, Holzman LB, Kopp JB.; J Am Soc Nephrol (2003 Aug) 14(8):1998-2003. Abstract/Full Text

Use of nitrite salts for the treatment of cardiovascular conditions (U.S. Patent 9, 387,224)

Fri, 06 Feb 2026 12:34:18 -0500

It has been surprisingly discovered that administration of nitrite to subjects causes a reduction in blood pressure and an increase in blood flow to tissues. The effect is particularly beneficial, for example, to tissues in regions of low oxygen tension. This discovery provides useful treatments to regulate a subject's blood pressure and blood flow, for example, by the administration of nitrite salts. Provided herein are methods of administering a pharmaceutically-acceptable nitrite salt to a subject, for treating, preventing or ameliorating a condition selected from:

ischemia-reperfusion injury (e.g., hepatic or cardiac or brain ischemia-reperfusion injury);
pulmonary hypertension (e.g., neonatal pulmonary hypertension); or
cerebral artery vasospasm.

Simple, rapid & reliable method for detecting thalassemia (U.S. Patent Number 5,281,519)

Fri, 06 Feb 2026 12:32:14 -0500

A simple, rapid and reliable method for diagnosis of thalassemia is described. The method comprises amplification of the cDNA by polymerase chain reaction and determining the ratio between α and β hemoglobin chain mRNAs.

Body Weight Simulator - Java Applet for Modeling Human Metabolism & Energy Expenditure for Adaptive Dieting & Exercise Regimens

Thu, 01 May 2025 09:10:34 -0400

Known methods for predicting weight loss fail to account for slowing of metabolism as weight is lost and therefore overestimate the degree of weight loss. While this limitation of the 3500 Calorie per pound rule has been known for some time, it was not clear how to dynamically account for the metabolic slowing. The invention provides a Java applet for modeling of human metabolism to improve the weight change predictions. The model has been validated using previously published human data and the model equations have been published. A web-based implementation of the published dynamic model has been created to allow users to perform simulations for planning weight loss interventions in adults and accounts for individual differences in metabolism and body composition. Values to the user include being able to see whether the target weight loss is realistic when the necessary caloric restriction, exercise and timeframe components are highlighted. Unrealistic goals become apparent. More moderate goals may result in greater long term success. The model also defines the caloric intake to be followed once the new weight target has been met to prevent weight being regained.

Personalized Body Weight Management System Using Monitoring Devices & Mathematical Models of Metabolism

Thu, 01 May 2025 09:09:47 -0400

Attempts to manage body weight are often unsuccessful or only temporary. This is, in part, due to antiquated dieting methods that attempt to address calorie consumption while ignoring metabolic and physical changes. Personalized and more comprehensive methods to track and manage body weight may be more effective. To that end, scientists at the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) developed and launched the Body Weight Planner that uses validated mathematical models of human metabolism to set weight management goals and predict individual body weight outcomes in the context of changing metabolic needs and calorie consumption.

More recently, developers at NIDDK have created a prototype personalized body weight management system prototype that builds on the science behind Body Weight Planner with the addition of patented tracking and feedback technology. This new system is targeted for use by professionals and is designed to be integrated into a comprehensive healthcare or wellness program. Improvements enable users to more accurately plan, track, and update personalized weight management interventions by accounting for changes in human appetite, metabolism, and calorie expenditure over time. There are opportunities for the prototype to be combined with other devices to provide data input through wearables and at-home measurements. This system provides meaningful feedback through enhanced functionality and features to meet weight management goals.

Therapeutic Polypeptides & Their Use (U.S. Patent Application Number 61/890,585)

Wed, 30 Apr 2025 11:06:44 -0400

NIDDK investigators have developed a synthetic polypeptide with beneficial effects and prospective treatment of chronic kidney disease (CKD). End stage renal disease (ESRD) is a major health and economic burden and requires transplantation or dialysis to avoid imminent death. ESRD is the final stage of CKD, which progresses over the course of decades. The major causes of CKD are diabetes and hypertension, and treatments for diabetes and hypertension are only partially effective to slow the progression of CKD. Only one class of drugs Angiotensin Converting Enzyme inhibitors and/or Angiotensin Receptor Blockers (ACEi/ARB) is effective for slowing CKD progression, but many CKD patients, especially African Americans, are resistant to ACEi/ARB therapy. Several anti-oxidant/anti-fibrotic agents (including bardoxolone and pirfenidone) have not shown efficacy in recent clinical trials. Therefore, novel therapeutic approaches are needed for this large unmet medical need. The investigators previously demonstrated in a mouse preclinical model that CD36 is important for developing CKD, where deletion of the CD36 gene prevented most of the progression of CKD. Subsequently, they tested whether peptides designed to inhibit CD36 action could also prevent the progression of CKD.

CKD is highly prevalent in the US, affecting more than 20 million people. African Americans have a higher risk than Caucasians.

UTX LoxP Mouse

Wed, 30 Apr 2025 10:55:20 -0400

UTX-flox. Conditional knockout mice for the histone demethylase UTX (Kdm6a) conditional knockout will help understand its role as a tumor suppressor.

Di- and tri-methylations on histone H3 lysine 27 (H3K27me2 and H3K27me3) are epigenetic marks for gene repression. UTX (ubiquitously transcribed X chromosome protein), also known as Kdm6a (lysine (K)-specific demethylase 6a) is a histone demethylase that specifically removes H3K27me2 and H3K27me3. UTX knockout mice are embryonic lethal, so we have generated UTX conditional knockout mice (UTX-flox) in which exon 24 is flanked with loxP sites. UTX has been found to be a tumor suppressor gene mutated in a wide variety of human cancers. The UTX-flox mice provide a valuable tool to study how UTX functions as a tumor suppressor and as an epigenetic regulator of gene expression.

Transframe peptide inhibitor of viral protease (U.S. Patent Number 5,872,210)

Wed, 30 Apr 2025 10:54:04 -0400

The present invention describes small, water soluble peptides isolated from a native virus inhibitory sequence that blocks maturation of the virally encoded protease and inhibits the mature protease as well. The peptides may be used in the treatment of virally infected cells, in the preparation of vaccine formulations, in the generation of clinically relevant antibodies and anti-idiotypic antibodies and in the generation of a screening assay or kit that can be used to identify other similarly acting protease inhibitors.

Tg(Wap-cre)11738Mam Mouse

Wed, 30 Apr 2025 10:41:08 -0400

Cre-recombinase under the control of the whey acidic acid protein was only detected in alveolar epithelial epithelial cells of mammary tissue during lactation, and transcription occurred at all stages of mammary development.

The Cre recombinase from bacteriophage P1 excises intervening DNA sequences located between two unidirectional lox sites positioned on the same linear DNA segment, leaving one lox site behind. Through insertion of lox sites via homologous recombination into the gene of interest and targeting Cre recombinase expression to a specific cell type using a tissue-specific promoter, it is possible to introduce predetermined deletions into the mammalian genome. To delete genes specifically from mammary gland, transgenic mice were created carrying the Cre gene under the control of the whey acidic protein (WAP) gene promoter. Expression of WAP-Cre was only detected in alveolar epithelial cells of mammary tissue during lactation. Recombination mediated by Cre under control of the WAP gene promoter was largely restricted to the mammary gland but occasionally was observed in the brain. High-level transcriptional activity of WAP-based transgenes can be obtained at every stage of mammary development.

Publication

Cre-mediated gene deletion in the mammary gland.: Wagner KU, Wall RJ, St-Onge L, Gruss P, Wynshaw-Boris A, Garrett L, Li M, Furth PA, Hennighausen L.; Nucleic Acids Res (1997 Nov) 25(21):4323-30. Abstract/Full Text

Tg(MMTV-Cre)#Mam Mouse

Wed, 30 Apr 2025 10:35:54 -0400

Cre-recombinase under the control of mouse mammary tumor virus long terminal repeat (MMTV) was expressed in the salivary gland and mammary epithelial cells of adult mice, and induced recombination in all tissues.

The Cre recombinase from bacteriophage P1 excises intervening DNA sequences located between two unidirectional lox sites positioned on the same linear DNA segment, leaving one lox site behind. Through insertion of lox sites via homologous recombination into the gene of interest and targeting Cre recombinase expression to a specific cell type using a tissue-specific promoter, it is possible to introduce predetermined deletions into the mammalian genome. To delete genes specifically from mammary gland, transgenic mice were created carrying the Cre gene under the control of the mouse mammary tumor virus (MMTV) long terminal repeat (LTR). In adult MMTV-Cre mice, expression of the transgene was confined to striated ductal cells of the salivary gland and mammary epithelial cells in virgin and lactating mice. In contrast to WAP-Cre, however, Cre expression under control of the MMTV LR resulted in recombination in all tissues.

Publication

Cre-mediated gene deletion in the mammary gland.: Wagner KU, Wall RJ, St-Onge L, Gruss P, Wynshaw-Boris A, Garrett L, Li M, Furth PA, Hennighausen L.; Nucleic Acids Res (1997 Nov) 25(21):4323-30. Abstract/Full Text

Tay-Sachs Disease Mouse

Wed, 30 Apr 2025 10:35:00 -0400

Also known as: Hexa^tm1Rlp, Hexa KO mouse

These Hexa knock-out mice exhibit accumulation of G_M2 ganglioside in the central nervous system. They are suitable for use in applications related to the study of Tay-Sachs disease in humans. Learn more on PubMed.

Mice can be obtained from Jackson Labs.

Synthesis & purification of hepatitis C virus-like particles (U.S. Patent Number 6,387,662)

Wed, 30 Apr 2025 10:34:35 -0400

Production of enveloped RNA virus-like particles intracellularly in vitro in insect cells using a recombinant baculovirus vector containing a cDNA coding for viral structural proteins is disclosed. In vitro production and purification of hepatitis C virus (HCV)-like particles containing HCV core protein, E1 protein and E2 protein is disclosed. Production of antibodies in vivo to the purified HCV-like particles is disclosed.

Stat5a LoxP/Stat5b LoxP (Stat5a/Stat5b tm2Mam) Mouse

Wed, 30 Apr 2025 10:32:00 -0400

Conditional knockout of Stat5a and Stat5b: Combined deletion of conserved Stat5a and Stat5b in mammary epithelium at different times during pregnancy reveal multiple distinct functions.

The signal transducer and activator of transcription (STAT) family of transcription factors conveys signals from membrane receptors to the nucleus, where they activate diverse genetic programs. Stat5a and Stat5b are highly conserved proteins that are activated by many cytokines, erythropoietin, prolactin and growth hormone. Despite their similarities, they have many unique functions. Stat5a deficiency results in the loss of prolactin-dependent mammary gland development, but does not affect body growth. Inactivation of Stat5b does not adversely affect mammary development and function, but leads to severe growth retardation. To study the effects of combined deficiency of Stat 5a and 5b before and during pregnancy, loxP was added to the ends of a DNA fragment that contains the two genes which are located within a stretch of 110 kb on chromosome 11 in a head to head orientation with no other genes between them. The loxP-flanked fragment was introduced into the genome using homologous recombination, and deleted using two transgenic lines expressing Cre in mammary epithelium at different times. Deletion of Stat 5 before pregnancy prevents epithelial proliferation. Ductal characteristics are retained but differentiation into secretory alveoli does not occur. When deletion of Stat5 occurs late in pregnancy after differentiation has started, differentiation is halted and premature death occurs.

Publication

Inactivation of Stat5 in mouse mammary epithelium during pregnancy reveals distinct functions in cell proliferation, survival, and differentiation.: Cui Y, Riedlinger G, Miyoshi K, Tang W, Li C, Deng CX, Robinson GW, Hennighausen L.; Mol Cell Biol (2004 Sep) 24(18):8037-47. Abstract/Full Text

Stat5a KO(Stat5a tm1Mam) Mouse

Wed, 30 Apr 2025 10:22:58 -0400

Stat 5a Knockout: Stat5a deficiency results in the loss of prolactin-dependent mammary gland development and lactogenesis.

Prolactin induces mammary gland development and lactogenesis. Binding of Prolactin to its receptor leads to the phosphorylation and activation of STAT (signal transducers and activators of transcription) proteins. Two Stat proteins, Stat 5a and Stat5b, are expressed in mammary tissues during pregnancy. Stat5a null mice developed normally, and were indistinguishable from hemizygous and wild-type littermates in size, weight and fertility. Mammary lobulo-alveolar outgrowth during pregnancy was reduced and females failed to lactate after parturition. Stat5b, despite 96% similarity to Stat5a, could not compensate for the loss of Stat5a. Stat5a is the principal and obligate mediator of mammopoietic and lactogenic signaling.

Publication

Stat5a is mandatory for adult mammary gland development and lactogenesis.: Liu X, Robinson GW, Wagner KU, Garrett L, Wynshaw-Boris A, Hennighausen L.; Genes Dev (1997 Jan) 11(2):179-86. Abstract/Full Text

Stat1LoxP (Stat1 tm1Mam) Mouse

Wed, 30 Apr 2025 10:21:03 -0400

Selective inactivation of Stat1 in mammary cells indicates that its effect as a tumor suppressor in breast is direct.

STAT1 is considered a tumor suppressor, but it is not known if this effect occurs directly in mammary cells or secondarily by disrupting interferon signaling through the JAK/STAT1 pathway to induce immune responses. ERBB2/neu-induced breast cancer appeared sooner in mice lacking STAT1 only in mammary cells than in wild-type mice, indicating that STAT1 tumor suppression was intrinsic to mammary cells and not secondary to an induced immune response.

Publication

Loss of STAT1 from mouse mammary epithelium results in an increased Neu-induced tumor burden.: Klover PJ, Muller WJ, Robinson GW, Pfeiffer RM, Yamaji D, Hennighausen L.; Neoplasia (2010 Nov) 12(11):899-905. Abstract/Full Text

Sphk2 KO Mouse

Wed, 30 Apr 2025 10:20:20 -0400

Also known as: Sphk2^tm1Rlp, sphingosine kinase 2 KO mouse

Sphingosine kinase 1 and 2, expressed by the Sphk1 and Sphk2 genes, respectively, are enzymes that catalyzes the synthesis of sphingosine-1-phosphate (S1P), a lipid signaling molecule, from sphingosine and ATP. S1P regulates immune cell trafficking and vascular function by signaling through the S1P family of GPCRs. The Sphk2^tm1Rlp allele expresses β-galactosidase (lacZ) from a disrupted Sphk2 gene. Mice homozygous for this allele are viable and fertile. Learn more on PubMed.

Mice can be obtained from Jackson Labs.

Sphk1 KO Mouse

Wed, 30 Apr 2025 10:20:05 -0400

Also known as: Sphk1^tm1Rlp, sphingosine kinase 1 KO mouse

Sphingosine kinase 1 and 2, expressed by the Sphk1 and Sphk2 genes, respectively, are enzymes that catalyzes the synthesis of sphingosine-1-phosphate (S1P), a lipid signaling molecule, from sphingosine and ATP. S1P regulates immune cell trafficking and vascular function by signaling through the S1P family of GPCRs. In the Sphk1^tm1Rlp allele , a neo cassette replaces exons 3-5 and part of exon 6, encoding the lipid kinase catalytic domain and putative nucleotide binding motif, of the sphingosine kinase 1 (Sphk1) gene, abolishing gene function. Mice homozygous for this allele are viable and fertile. Learn more on PubMed.

Mice can be obtained from Jackson Labs.

Method for Spectrally Selective B1 Insensitive T2 Preparation Contrast Enhancement for High Field Magnetic Resonance Imaging

Wed, 30 Apr 2025 10:19:06 -0400

A T₂ preparation sequence uses a segmented BIR-4 adiabatic pulse with two substantially equal delays and is insensitive to B₁ field variations and can simultaneously suppress fat signals with low specific absorption rate (SAR). An adiabatic reverse half passage pulse is applied followed by a predetermined delay. An adiabatic full passage pulse is applied followed by a substantially equal delay, followed by an adiabatic half passage pulse. Fat signal suppression is achieved by increasing or decreasing either the first delay or the second delay.

Small volume viscometer

Wed, 30 Apr 2025 10:16:16 -0400

NIDDK scientists have developed a capillary-based device and system for measuring the rheological properties of solutions of synthetic and biological polymers. The device automatically serially dilutes and varies the flow rate of a sample, permitting measurement of solution viscosity across wide ranges of concentration and shear rate without changing samples. Viscosity is calculated directly from Poiseuille's law, given the measured difference in pressure between two ends of a capillary tube through which the solution is flowing at a known rate. The device can rapidly and accurately assay solute stability, solute-solvent and solute-solute interactions in solutions of proteins and other macromolecules of biotechnological and pharmaceutical interest, as well as solution injectability.

Single electrode corona discharge electrochemical/electrospray ionization (U.S. Patent Number 7,759,643)

Wed, 30 Apr 2025 10:12:42 -0400

A single electrode electrochemical/electrospray ionization source using a corona discharge and a method of analyzing a sample using a corona discharge single electrode electrochemical/electrospray ionization source are provided. In the corona discharge single electrode electrochemical/electrospray ionization technique electrons are removed from the metal tip of the device through gases present in the electrospray ion source resulting in electrochemical ionization of the sample of interest. The resulting odd electron sample cation (positive ion mode) or anion (negative ion mode) can then be analyzed by an appropriate technique, such as, for example, a mass spectrometer.

Sgpp2 KO Mouse

Wed, 30 Apr 2025 10:09:59 -0400

Also known as: Sgpp2^tm1Rlp, sphingosine-1-phosphate (S1P) phosphatase 2 KO mouse

Intracellular metabolism of sphingosine-1-phosphate (S1P) is controlled, in part, by two homologous S1P phosphatases (SPPases), 1 and 2, which are encoded by the Sgpp1 and Sgpp2 genes, respectively. SPPase activity is needed for efficient recycling of sphingosine into the sphingolipid synthesis pathway. Sgpp2 KO mice exhibit decreased adaptive pancreatic β-cell proliferation and β-cell endoplasmic reticulum stress, revealing the importance of the sphingolipid recycling pathway in β-cell physiology. Learn more on PubMed.

Mice can be obtained by contacting the NIDDK Technology Advancement Office.

Sandhoff Disease Mouse

Wed, 30 Apr 2025 10:09:47 -0400

Also known as: Hexb^tm1Rlp, Hexb KO mouse

Hexb KO mice develop motor defects beginning at about 3 months of age. The defects progressively worsen and homozygous mice die by 4.5 months of age. Mice display gangliosidosis; mice abnormally accumulate GM2 and GA2 ganglioside and serve as a model of Sandhoff disease. Learn more on PubMed.

Mice can be obtained from Jackson Labs.

S1pr3 KO Mouse

Wed, 30 Apr 2025 10:09:17 -0400

Also known as: S1pr3^tm1Rlp, sphingosine-1-phosphate receptor 3 KO mouse, S1P3 KO mouse, Edg3 KO mouse

S1pr3 is a member of the sphingosine-1-phosphate G-protein coupled receptor family. S1pr3 KO mice are grossly normal. Double KO S1pr2/S1pr3 mice exhibit incompletely penetrant lethality as embryos. Learn more on PubMed.

Mice can be obtained from MMRRC.

S1pr2 KO Mouse

Wed, 30 Apr 2025 10:09:00 -0400

Also known as: S1pr2^tm1Rlp, sphingosine-1-phosphate receptor 2 KO mouse, S1P2 KO mouse, Edg5 KO mouse

S1pr2 is a member of the sphingosine-1-phosphate G-protein coupled receptor family. S1pr2 KO mice exhibit deafness, seizures and are prone to B-cell lymphoma development. Double KO S1pr2/S1pr3 mice exhibit incompletely penetrant lethality as embryos. Learn more on PubMed.

Mice can be obtained from MMRRC.

Floxed S1pr1 Mouse

Wed, 30 Apr 2025 10:07:34 -0400

Also known as: S1pr1^tm2.1Rlp, floxed S1P1 mouse

S1pr1 is a member of the sphingosine-1-phosphate G-protein coupled receptor family. S1pr1 is essential for vascular maturation during embryonic development and is involved in in the regulation of innate and adaptive immune responses by controlling lymphocyte egress from the thymus, spleen, bone marrow, and lymph nodes. It has also been implicated in the regulation of vascular function. These S1pr1^loxP/loxP mice possess loxP sites flanking exon 2 of the sphingosine-1-phosphate receptor 1 (S1pr1) gene. Mice that are homozygous for the floxed allele are viable and fertile. When these mutant mice are bred to mice that express Cre recombinase, resulting offspring will have S1pr1 deleted in cre-expressing tissues. Learn more on PubMed.

Mice can be obtained from Jackson Labs.

S1pr1 KO Mouse

Wed, 30 Apr 2025 10:06:22 -0400

Also known as: S1pr1^tm1Rlp, sphingosine-1-phosphate receptor 1 KO mouse, S1P1 KO mouse, Edg1 KO mouse

S1pr1 is a member of the sphingosine-1-phosphate G-protein coupled receptor family. S1pr1 is essential for vascular maturation during embryonic development and is also involved in cell survival, migration, adhesion, and proliferation. This receptor plays a role in the regulation of innate and adaptive immune responses by controlling lymphocyte egress from the thymus, spleen, bone marrow, and lymph nodes. It has also been implicated in the regulation of vascular function. S1pr1 KO mice exhibit embryonic hemorrhage leading to intrauterine death between E12.5 and E14.5. Learn more on PubMed.

Mice can be obtained from MMRRC.

S1P1 GFP Signaling Mouse

Wed, 30 Apr 2025 10:06:05 -0400

Also known as: S1pr1^{tm3.1(tTA,-Arrb2)Rlp}, S1P1 Tango mouse

These S1pr1 knockin mice have a bicistronic transcription unit inserted at the C-terminus of exon 2 of the sphingosine-1-phosphate receptor 1 (S1pr1) gene. Specifically, the allele contains two fusion proteins, a tetracycline-regulated transactivator (tTA)-tobacco etch virus (TEV) protease recognition sequence (tevs) fusion protein and an Arrb2 (murine β-arrestin)-TEV protease fusion protein, separated by an internal ribosome entry site (IRES). Homozygous mice are viable and fertile. S1PR1 is a G protein-coupled receptor (GPCR) that acts as a regulator of vascular development and function in endothelial cells, and has been shown to play a role in autoimmunity and inflammation. In this strain, S1PR tethers tTA to the membrane. Ligand activation of S1PR1 results in phosphorylation of the receptor and subsequent recognition of the tevs sequence by the β-arrestin-TEV protease fusion protein, triggering the release of tTA. Free tTA is then able to enter the nucleus where it regulates transcription of genes of interest under the regulatory control of a tetracycline-responsive promoter element (TRE; tetO). The transcriptional regulation elicited by tTA is blocked by the administration of the tetracycline analog, doxycycline.

When mated to Tg(tetO-HIST1H2BJ/GFP)47Efu/J transgenic mice (Jackson Lab: Stock No. 005104), resulting S1P1 GFP signaling mice exhibit tTA-induced dox-dependent green fluorescent protein expression in cells where S1P1 signaling pathway is activated. Fluorescence is evident in thymi, spleen, lymph nodes, lungs, and heart. Learn more on PubMed.

Mice can be obtained from Jackson Labs.

S1P1 Luciferase Signaling Mouse

Wed, 30 Apr 2025 10:05:28 -0400

S1P₁ luciferase signaling mice enable the bioluminescent detection of sphingosine-1-phosphate receptor 1 (S1P₁) activation in real time. A synthetic S1P₁ signaling pathway, designed to report the interaction between S1P₁ and β-arrestin2 via the firefly split luciferase fragment complementation system, is genetically encoded in these mice. Upon receptor activation and subsequent β-arrestin2 recruitment, an active luciferase enzyme complex is produced, which can be detected by in vivo bioluminescence imaging. Learn more on PubMed.

Mice can be obtained by contacting the NIDDK Technology Advancement Office.

Purine Derivatives as A3 Receptor-selective Agonists (U.S. Patent Application Number 13/371,081)

Wed, 30 Apr 2025 09:58:34 -0400

Disclosed are (N)-methanocarba adenine nucleosides, e.g., of formula (I) as highly potent A3 adenosine receptor agonists, pharmaceutical compositions comprising such nucleosides, and a method of use of these nucleosides, wherein R1-R6 are as defined in the specification. These nucleosides exhibit similar selectivities as agonists of the A3 versus the A1 receptor for both human and mouse adenosine receptors, and are contemplated for use in the treatment a number of diseases, for example, inflammation, cardiac ischemia, stroke, asthma, diabetes, and cardiac arrhythmias.

Methods of preventing viral replication (U.S. Patent Number 5,610,050)

Wed, 30 Apr 2025 09:51:06 -0400

The invention relates to methods and compositions for inhibition of viral replication. In particular, termination of replication of hepatitis B virus is achieved by introducing into a target cell an antisense oligonucleotide having a sequence substantially complementary to an mRNA which is in turn complementary to a portion of the minus strand of a hepatitis viral genome, which portion encoding solely part or all of the terminal protein region of the viral polymerase.

Potent Nucleotide Inhibitors of Ecto-5'-Nucleotidase (CD73)

Wed, 30 Apr 2025 09:49:49 -0400

Novel, small molecule nucleotide derivatives have been developed containing either a purine or pyrimidine nucleobase that competitively block the enzyme CD73, or ecto-5'-nucleotidase. CD73 converts extracellular AMP to adenosine, the native activator of 4 subtypes of adenosine receptors. CD73 has been shown to be upregulated around tumors leading to excess production of adenosine. The use of CD73 inhibitors in conjunction with cancer immunotherapy is being explored in preclinical research and clinical trials. Blocking this enzyme can greatly reduce immunosuppressive adenosine in the tumor microenvironment. Therefore, combined therapy is expected to be more effective than cancer immunotherapy alone.

Previously, adenine nucleotides have been used as CD73 inhibitors in research, but these can lead to off-target activity at purinergic receptors. Thus, the ability to potently inhibit the enzyme using a different nucleobase derivative can provide greater specificity for the target in comparison to earlier inhibitors.

Polycystic kidney disease gene (U.S. Patent Number 5,654,170)

Wed, 30 Apr 2025 09:49:08 -0400

The present invention involves isolated nucleic acid encoding human PKD1, and sequences derived therefrom. The invention also encompasses vectors comprising these nucleic acids, host cells transformed with the vectors, and methods for producing PKD1 protein or fragments thereof. In another aspect, the invention involves isolated oligonucleotides that hybridize only to the authentic expressed PKD1 gene, and not to PKD1 homologues. In yet another aspect, the invention involves isolated mutant PKD1 genes, and their cDNA cognates. Further provided are isolated oligonucleotides that discriminate between normal and mutant versions of the PKD1 gene. Methods and compositions for treating APKD or disease conditions having the characteristics of APKD are also provided.

Polycystic kidney disease gene & protein (U.S. Patent Number 6,071,717)

Wed, 30 Apr 2025 09:48:29 -0400

Polycystic kidney disease gene (Patent Number 6,867,288)

Wed, 30 Apr 2025 09:47:53 -0400

The present invention provides methods and compositions for treating cyst formation in PKD1-associated epithelial cells. Such methods encompass administering an isolated human PKD1 gene, or fragments of the gene, under conditions that result in expression of therapeutically effective amounts of all, or part of, the PKD1 protein. The invention also encompasses compositions for treating cyst formation associated with APKD.

PNA-Detection of Pathogens (U.S. Patent Number 13/592,490)

Wed, 30 Apr 2025 09:38:17 -0400

The laboratory of Daniel Appella, PhD. has developed a prototype HIV testing kit that employs a peptide nucleic acid probe to capture HIV RNA recovered from patient blood. The probe targets a highly conserved segment of the Gag sequence. Initial data from tests run side by side with PCR show high sensitivity and specificity. The test, excluding the cost for an RNA extraction kit, has been calculated to be under $0.70 per sample based on retail price lists for reagents. This is within the cost target suggested by the World Health Organization. It is hoped that the test would be suitable for use in a developing country. While the test has not been optimized, operated as is, the assay would take 3-4 hours providing results while a patient is still at the clinic.

Pkd mutations & evaluation of same (U.S. Patent Number 10,760,128)

Wed, 23 Apr 2025 15:16:13 -0400

The present invention relates to methods of detecting novel mutations in a PKD1 and/or PKD2 gene that have been determined to be associated with autosomal dominant polycystic kidney disease (ADPKD) in order to detect or predict the occurrence of ADPKD in an individual.

Method for over-expression of rapid purification of biosynthetic proteins (U.S. Patent Number 6,077,694)

Wed, 23 Apr 2025 15:14:42 -0400

The subject invention relates to a method of producing and purifying large quantities of a biosynthetic protein. The gene which codes for the protease is placed between the binding domain of a gene which codes for a binding protein and a gene coding for the target protein of interest. The fused gene construct is inserted in an expression vector which is then introduced into a host cell.

Detection of nucleic acid sequence variations using phase Mu transposase (U.S. Patent Number 7,316,903)

Wed, 23 Apr 2025 15:12:43 -0400

The present invention relates, e.g., to a method of detecting a mismatch in a double stranded nucleic acid target, comprising (a) contacting the target with (i) a Mu-end nucleic acid, and (ii) a phage Mu transposase, under conditions effective for the Mu-end nucleic acid to transpose into the target at about the site of a mismatch, if the target comprises a mismatch, and (b) detecting transposition of the Mu-end DNA into the target, wherein transposition of the Mu-end nucleic acid into the target at a predominant site indicates the presence of a mismatch at that site.

Novel conjugate vaccine process, cholera vaccine (U.S. Patent Number 61/507,054)

Wed, 23 Apr 2025 15:11:40 -0400

A new conjugate vaccine for cholera has been developed. The invention includes a new method to conjugate the O-specific polysaccharide-core part of the bacterial lipopolysaccharide and protein subcomponents. Conventional technology has entailed chemical treatment of both components to introduce linkers, which made them amenable for covalent linking. The new method simplifies production by utilizing squaric acid chemistry for conjugating the free amine-containing species (e.g., polysaccharides) directly to amine-containing species (e.g., proteins) without prior modification of either component. While demonstrated in this new cholera prototype vaccine, the technology is envisioned as generally applicable, thereby streamlining a complex production process.

Programmable self-assembled nanostructures based on sidechain-modified pna for the multivalent display of ligands (U.S. Patent Number EP 2569328 A2)

Wed, 23 Apr 2025 15:10:06 -0400

The invention concerns compositions comprising strands of polynucleotide and strands of PNA, each PNA strand comprising:

(i) from 2 to 50 nucleobase subunits and
(ii) one or more gamma substituents.

The PNA strands are complementary to at least a portion of at least some of the polynucleotide strands, and the molar ratio of PNA strands to polynucleotide strands being at least 1:1. Certain gamma substituents are capable of effecting attachment of a PNA strand to a cell. The invention also concerns construction of nanostructure platforms and vaccines and use of the inventive compositions in inhibiting disease states in mammals.

Mutant Mouse: Kmt2d (lysine (K)-specific methyltransferase 2D) (MII4 LoxP Mouse)

Wed, 23 Apr 2025 15:02:55 -0400

Mll4-flox. Conditional knockout mice for the histone methyltransferase Mll4 (Kmt2d) will help understand its role as a tumor suppressor.

Mono- and di-methylations on histone H3 lysine 4 (H3K4me1 and H3K4me2) are epigenetic marks for transcriptional enhancers, which control cell type-specific gene expression. Mll4, also known as Kmt2d (lysine (K)-specific methyltransferase 2D), is a major mammalian histone H3K4 mono- and di-methyltransferase enriched on enhancers (http://www.ncbi.nlm.nih.gov/pubmed/24368734). Mll4 knockout mice are early embryonic lethal, so we have generated Mll4 conditional knockout mice (Mll4-flox) in which exons 16-19 are flanked with loxP sites. Mll4 has been found to be a tumor suppressor gene mutated in a wide variety of human cancers. The Mll4-flox mice provide a valuable tool to study how Mll4 functions as a tumor suppressor and as an epigenetic regulator of cell-type-specific gene expression, cell differentiation and mouse development.

Publication

H3K4 mono- and di-methyltransferase MLL4 is required for enhancer activation during cell differentiation.: Lee JE, Wang C, Xu S, Cho YW, Wang L, Feng X, Baldridge A, Sartorelli V, Zhuang L, Peng W, Ge K.; Elife (2013 Dec) 2:e01503. Abstract/Full Text

Mutant Mouse: Kdm6a (lysine (K)-specific demethylase 6A) (UTX Knockin Mouse)

Wed, 23 Apr 2025 14:52:38 -0400

UTX-KI. Enzyme-dead knockin (KI) mice for the histone demethylase UTX (Kdm6a) will help understand its role in tumor suppression and stem cell-mediated tissue regeneration.

Di- and tri-methylations on histone H3 lysine 27 (H3K27me2 and H3K27me3) are epigenetic marks for gene repression. UTX (ubiquitously transcribed X chromosome protein), also known as Kdm6a (lysine (K)-specific demethylase 6a), is a histone demethylase that specifically removes H3K27me2 and H3K27me3. To investigate the physiological role of UTX enzymatic activity, we have generated UTX enzyme-dead knockin mice (UTX-KI) which possess the H1146A and E1148A point mutations in exon 24 (http://www.ncbi.nlm.nih.gov/pubmed/26999603). UTX-KI mice are viable and fertile but show defects in muscle regeneration. UTX has been found to be a tumor suppressor gene mutated in a wide variety of human cancers. The UTX-KI mice provide a valuable tool to study how UTX functions as a tumor suppressor and as an epigenetic regulator of stem cell-mediated tissue regeneration.

Publication

UTX demethylase activity is required for satellite cell-mediated muscle regeneration.: Faralli H, Wang C, Nakka K, Benyoucef A, Sebastian S, Zhuang L, Chu A, Palii CG, Liu C, Camellato B, Brand M, Ge K, Dilworth FJ.; J Clin Invest (2016 Apr) 126(4):1555-65. Abstract/Full Text

MUP-tTA Mouse

Wed, 23 Apr 2025 14:48:25 -0400

Tetracycline-responsive transcriptional activator driven by the liver-specific mouse major urinary protein promoter (MUP-tTA).

The E. Coli tetracycline operon regulatory system was used to generate a liver-specific transcription activation system that was inhibited by tetracycline. The transcription activator was a fused protein consisting of a tetracycline repressor gene (tetR) that was only active in the presence of tetracycline and a herpes simplex virus protein (VP-16) transcription activating domain (Tet-Off). Transcription was induced only in the absence of tetracycline (Tet-Off). A liver-specific promoter such as the mouse major urinary protein (MUP) promoter determined that the tetracycline-regulated transcriptional activator (tTA) would be expressed specifically in liver. To study the effect of the transcription activator on a target gene (for example, beta-galactosidase, LacZ) specifically in liver, MUP-tTA mice would be mated with transgenic mice in which the TAg Target gene was controlled by the E.Coli Tetracycline Operator (Tet-O).

Publication

Conditional liver-specific expression of simian virus 40 T antigen leads to regulatable development of hepatic neoplasm in transgenic mice.: Manickan E, Satoi J, Wang TC, Liang TJ.; J Biol Chem (2001 Apr) 276(17):13989-94. Abstract/Full Text

Mini Mu containing plasmid & a method for rapid DNA sequencing (U.S. Patent Number 4,716,105)

Wed, 23 Apr 2025 14:46:14 -0400

The present invention discloses a rapid method of sequencing a relatively large segment of deoxyribonucleic acid. The method in part comprises high frequency insertion of a suitable transposon into a segment of DNA of interest. Preferable use of Mu transposons is described. A plasmid having mini-Mu transposons has been prepared and disclosed.

Mice with Natural Gastrointestinal Microbiota

Wed, 23 Apr 2025 14:28:40 -0400

Among 21 distinct mouse populations worldwide, we identified a closely related wild relative to standard laboratory mouse strains. Its bacterial gut microbiome differed significantly from its laboratory mouse counterpart and was transferred to and maintained in laboratory mice over several generations. Laboratory mice reconstituted with natural microbiota exhibited reduced inflammation and increased survival following influenza virus infection and improved resistance against mutagen/inflammation-induced colorectal tumorigenesis. This model should enable the discovery of mechanisms that are relevant for host fitness and protection against diseases in the natural world and improve the modeling of complex diseases of free-living mammals.

Publication

Wild Mouse Gut Microbiota Promotes Host Fitness and Improves Disease Resistance.: Rosshart SP, Vassallo BG, Angeletti D, Hutchinson DS, Morgan AP, Takeda K, Hickman HD, McCulloch JA, Badger JH, Ajami NJ, Trinchieri G, Pardo-Manuel de Villena F, Yewdell JW, Rehermann B.; Cell (2017 Nov) 171(5):1015-1028. Abstract/Full Text

Methanocarba cycloalkyl Nucleoside Analogues (U.S. Patent Number 7,790,735)

Wed, 23 Apr 2025 14:24:20 -0400

The present invention provides novel nucleoside and nucleotide derivatives that are useful agonist or antagonists of P1 and P2 receptors. For example, the present invention provides a compound of formula A-M, wherein A is modified adenine or uracil and M is a constrained cycloalkyl group. The adenine or uracil is bonded to the constrained cycloalkyl group. The compounds of the present invention are useful in the treatment or prevention of various diseases including airway diseases (through A2B, A3, P2Y2 receptors), cancer (through A3, P2 receptors), cardiac arrhythmias (through A1 receptors), cardiac ischemia (through A1, A3 receptors), epilepsy (through A1, P2X receptors), and Huntington's Disease (through A2A receptors).

(N)-Methanocarba Adenosine Derivatives & Their Dendrimer Conjugates as A3 Receptor Agonists (U.S. Patent Application Number 61/313,961)

Wed, 23 Apr 2025 14:23:03 -0400

Disclosed are (N)-methanocarba adenine nucleosides, e.g., of the formula (I): as A₃ adenosine receptor agonists, pharmaceutical compositions comprising such nucleosides, and a method of use of these nucleosides, wherein A, a, R2, and R3 are as defined in the specification. These nucleosides are contemplated for use in the treatment a number of diseases, for example, inflammation, cardiac ischemia, stroke, asthma, diabetes, and cardiac arrhythmias. Also disclosed are conjugates comprising a dendrimer and one or more ligands, which are functionalized congeners of an agonist or antagonist of a receptor of the G-protein coupled receptor (GPCR) superfamily. Such conjugates are have the potential of being used as dual agonists, dual antagonists, or agonist/antagonist combinations.

mEpoR KO / Tg(hEpoR) Mouse

Wed, 23 Apr 2025 14:18:40 -0400

mEpoR-/- hEpoR+: The mouse Erythropoietin Receptor knockout that contains a human Erythropoietin Receptor transgene can be used to define the potency of recombinant erythropoietin preparations used to treat anemia associated with chronic kidney disease.

Erythropoietin, acting by binding to Erythropoietin receptors (EpoR) on erythroid progenitor cells, is required for erythropoiesis. Absence of erythropoietin or the EpoR in mice interrupts erythropoiesis in the fetal liver and result in death at embryonic day 13.5. An 80-kb human EpoR transgene bred onto a mouse EpoR null background (provided by F. Constantini of Columbia University) restored effective erythropoiesis in the EpoR null mouse. Erythropoietin preparations made utilizing recombinant DNA technology are used in the treatment of anemia in chronic kidney disease and other critical illnesses. The mouse EpoR null mouse containing the human EpoR transgene can be used to define the potency of erythropoietin preparation in humans.

Publication

The human erythropoietin receptor gene rescues erythropoiesis and developmental defects in the erythropoietin receptor null mouse.: Yu X, Lin CS, Costantini F, Noguchi CT.; Blood (2001 Jul) 98(2):475-7. Abstract/Full Text

MEN1, the gene associated with multiple endocrine neoplasia type 1, menin polypeptides & uses thereof (U.S. Patent Number 7,358,347)

Wed, 23 Apr 2025 14:16:45 -0400

The invention relates to the discovery of a novel tumor suppressor gene which is associated with multiple endocrine neoplasia type 1. The gene has been designated MEN1 and the gene product is menin. The absence of this protein and associated mutations in the corresponding gene have been identified in individuals suffering from multiple endocrine neoplasia type 1. The identification of this marker for multiple endocrine neoplasia type 1 has diagnostic uses as well as for gene therapy.