Several crosslinking strategies exist to generate injectable materials. However, the vast majority of materials have very rapid gelation kinetics, which do not allow for complex injection routes via catheter where the material must remain at body temperature inside the catheter for a period of time. An example of such a delivery route is cardiac catheter delivery, where the material must remain liquid in the catheter to facilitate multiple injections to the damaged region of the heart. The delivery of injectable materials into the heart has been studied to encourage endogenous cell infiltration and repair as well as for the delivery of cells or other therapeutics. These studies have, however, largely been performed in small animals since the gelation kinetics of most injectable materials prevent cardiac catheter delivery. (more...)

Chronic pruritus is estimated to occur in about 8% of the adult population. However, there are few drugs specifically targeting this problem. With a growing interest in this area, new drugs may be developed to address this problem. Screening active compounds using current methods, such as manual counting in real time or recorded videos, can be time consuming. Accordingly, there is a need to automate detection of scratching in test animals. (more...)

Many psychiatric drugs have serious side effects that are often due to general toxicity and a lack of specific drug action, therefore developing a drug more precisely against a gene or the pathway can potentially enable higher efficacy and less toxicity.Building on previous studies on a large Scottish schizophrenic family, UCSD researchers have delineated a novel pathway that can potentially enable more effective drug development. They found that the component genes of this pathway have cytotoxic effects and likely to contribute to the reduction of brain volume in schizophrenic patients. Therefore screening for drugs that inhibit the cytotoxicity brought on by these genes may reduce the pathogenesis of schizophrenia. (more...)

Bone metastases are detected in 80-100% of men who die of prostate cancer and such metastasis leads to painfully debilitating fractures, spinal compression and rapid decline. In addition, metastases to bone tissue often become resistant to standard therapies including androgen deprivation, radiation and chemotherapy. The scarcity of primary human prostate cancer bone metastasis tissues has crippled direct analysis and options to better understand the disease. In addition, spontaneous bone metastasis of prostate cancer in murine models, even to implanted human bone, is an exceedingly rare event. The development of a patient-derived xenograft (PDX) model of bone metastatic prostate cancer presents new options for improving therapeutic options. (more...)

HBV infection can lead to chronic infections that result in 0.6 million deaths per year worldwide by causing liver failure and cancer. Clearance of HBV infection is age dependent, with the majority of adult-acquired infections leading to spontaneous clearance, whereas infection in young children often leads to chronic infections. To study these early events of infection and immune activation that lead to HBV clearance or persistence, in vivo models are needed to screen and validate lead drug candidates. HBV cannot infect mice, however, researchers at UCSF have generated transgenic mouse models that mimic critical features of primary HBV infection observed in humans. (more...)

Although mammals have been cloned from genetically manipulated cultured cells, there is still a need for lower vertebrates to be used. Since zebrafish are important lower vertebrates to use for the study of vertebrate development, cloned zebrafish may offer an alternative to embryonic stem cells. Zebrafish cell cultures exhibit some characteristics of embryonic stem cells and are easier to use in research involving genetic manipulation. However, only short-term cell cultures of zebrafish exist for the study of vertebrate development. (more...)

A crucial limitation to our understanding of Alzheimer's disease (AD) is the inability to test hypotheses on live, patient-specific neurons. Patient autopsies are limited in supply and only reveal endpoints of disease. Rodent models harboring familial AD mutations lack important pathologies, and animal models have not been useful in modeling the sporadic form of AD because of complex genetics. The recent development of induced pluripotent stem cells (iPSCs) provides a method to create live, patient-specific models of disease and to investigate disease phenotypes in vitro. A proper understanding of the initiating events of AD and the existence of live disease models that accurately recapitulate the pathogenesis would lead to a much better informed therapeutic development effort. (more...)

RIN2 is a RAS effector protein with guanine nucleotide exchange factor (GEF) activity for RAB5 family GTPases. RIN2 is part of a cerebral cavernous malformations (CCM) signaling complex implicated in sporadic or inherited vascular lesions of the central nervous system. Additionally, RIN2 deficiencies are linked to the human connective tissue disorders MACS syndrome2 and RIN2 syndrome3. UCLA researchers have developed a novel polyclonal antibody to detect the human RIN2 protein. The efficacy of this antibody has been confirmed for a number of applications, including immunoprecipitation, immunoblot and immunofluorescence. (more...)

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A line of knockout mouse hase been developed that deletes the 2a subunit of the brain Na channel. This mouse model makes possible the modellingof a variety of neurologic disorders, drug target validation, and the screening of neuroactive drug candidates for efficacy, side effects and mechanism of action. (more...)

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Use of synthetic nucleic acids to manipulate gene function has become a powerful tool for both basic research and therapeutics.  Silencing disease targets by RNA interference is a promising approach to drug development, and various experimental RNA therapies are currently in clinical development by both small and large biotechnology companies.  miRNAs are also being developed for disease treatment and diagnosis.  However, lack of specifically targeted, efficient and safe vehicles for systemic delivery of small RNA payloads in vivo is a serious challenge.  Synthetic nucleic acids face a number of physiological barriers in the bloodstream, and their intracellular uptake is hampered by the fact that they are highly charged and have much larger molecular wieght than small-molecule drugs.  Current strategies to circumvent these problems includes local administration, chemical modifications of nucleic acids, viral delivery vectors, lipid-based delivery systems, polymer-based delivery systems and nanoparticle encapsulation.  These methods have serious flaws including toxicity, inummue effects, non-selectively and high cost of manufacturing.  Therefore, novel ways to deliver synthetic nucleic acids for use in humans and experimental animal models are sorely needed. (more...)

Huntington's disease (HD) is an adult-onset, autosomal-dominant neurodegenerative disease that is clinically characterized by a triad of movement disorders (i.e., chorea and bradykinesia), psychiatric symptoms, and cognitive deficits. In afflicted patients, symptoms usually progress relentlessly until death in 15-20 years after disease onset. HD is one of nine neurodegenerative disorders caused by a CAG repeat expansion encoding a polyglutamine (polyQ) repeat in otherwise unrelated proteins. In HD, the mutated Huntingtin (mhtt) protein is ubiquitously expressed in both neuronal and nonneuronal tissues. The polyQ repeat, located in the N terminus of huntingtin (htt), is normally less than 36, but is expanded to more than 37 in HD patients. In all polyQ disorders, there is an inverse relationship between the length of polyQ and the age of disease onset. Currently, there is no effective treatment or cure for HD or any other polyQ disorder. (more...)

Asthma, a chronic inflammatory disease of the lung, affects an increasingly larger population every year and presents a major public health problem in terms of morbidity and cost. The cause and mechanisms of asthma and allergic diseases are not yet understood. As the spontaneous development of asthma in a non-human animal is next to impossible, the ability to engineer mouse models of the disease remains critical to the understanding and treatment of the disease. (more...)

UCLA researchers have developed knockout mice in which the Synaptotagmin 4 (Syt4) gene is deleted. Synaptotagmin 4 is involved in a number of neurobiological events, including vesicle fusion and neurotransmitter release at neuronal synapses. These mice can be used to study many central nervous system (CNS) functions, including memory, learning, fear and anxiety (more...)

Prostaglandin-endoperoxide synthase 2 (Ptgs2, also known as Cyclooxygenase-2 or Cox-2) is an enzyme involved in the biosynthesis of prostanoids. Aberrant Ptgs2 expression plays major causal and modulatory roles in various cancers, neurodegenerative diseases, and chronic and acute inflammation conditions. UCLA researchers have developed a transgenic mouse model with an insertion of the firefly luciferase reporter gene into the endogenous Ptgs2 locus. This results in concomitant expression of Ptgs2 and the luciferase reporter, allowing researchers to non-invasively monitor Ptgs2 expression in vivo. (more...)

The ability to target gene deletion to a specific cellular compartment via the Cre/loxP system has been a powerful tool in the analysis of broadly expressed genes. Researchers at UCLA generated a transgenic mouse line in which expression of Cre-recombinase is under the regulatory control of the VE-Cadherin promoter. Temporal distribution and activity of the enzyme was evaluated with two independent Cre reporter lines. Histological analysis was performed throughout development and in the adult. Recombination of lox P sites with subsequent expression of beta-galactosidase or GFP was detected as early as E7.5 in endothelial cells of the yolk sac. Progressive staining of the embryonic vasculature was noted from E8.5-13.5; however, more contiguous reporter expression was only seen by E14.5 onward in all endothelial compartments including arteries, veins, and capillaries. In addition, the team found Cre activity in lymphatic endothelial cells. Unlike other endothelial-specific Cre mice, this model showed expression in the adult quiescent vasculature. Furthermore, the constitutive nature of the VE-Cadherin promoter in the adult can be advantageous for analysis of gene deletion in pathological settings. In addition, these mice may be useful in studies of the cardiovascular system, including angiogenesis, and endothelial and hematopoietic cell lineages. (more...)

To introduce temporal control in genetic experiments targeting the endothelium, we established a mouse line expressing tamoxifen-inducible Cre-recombinase (Cre-ERT2) under the regulation of the vascular endothelial cadherin promoter (VECad). Specificity and efficiency of Cre activity was documented by crossing VECad-Cre-ERT2 with the ROSA26R reporter mouse, in which a floxed-stop cassette has been placed upstream of the -galactosidase gene. We found that tamoxifen specifically induced widespread recombination in the endothelium of embryonic, neonatal, and adult tissues. Recombination was also documented in tumor-associated vascular beds and in postnatal angiogenesis assays. Furthermore, injection of tamoxifen in adult animals resulted in negligible excision (lower than 0.4%) in the hematopoietic lineage. The VECad-Cre-ERT2 mouse is likely to be a valuable tool to study the function of genes involved in vascular development, homeostasis, and in complex processes involving neoangiogenesis, such as tumor growth. (more...)

UCLA researchers have developed transgenic mice in which two exons of the Fox-2 (Rbfox2, RBM9) gene are flanked by loxP sites to permit deletion of the gene when the mice are crossed to mice expressing Cre recombinase. Fox-2 is an RNA-binding protein that regulates the alternative splicing of multiple transcripts in neurons and embryonic stem cells (ESCs). Fox-2 has been implicated in neurological disease, including spinocerebellar ataxia type I (SCAI), and has been shown to regulate the alternative splicing of ~7% of all genes in human ESCs. These mice are valuable research tools that can be used to analyze the development and differentiation of ESCs and brain in vivo. (more...)

UCLA investigators have characterized an association between the expression of a single gene and behavioral learning associated with forebrain function. The gene is preferentially expressed in the cell bodies and dendrites of post-natal neurons of the forebrain. Since the geneis not expressed until after birth, it does not appear to be involved in development. To assess the physiological role of the gene, knockout (genedisruption) mice were developed. The null mice appear normal and reproduce normally, but show enhanced amygdala-dependent long-term memory consolidation with a concomitant elevation in amygdala, but not hippocampal, long-term potentiation (LTP). Hippocampal-dependent learning and motorskills appear normal, but mutant mice showed specific enhancement ofamygdala-dependent learning of fear as assessed by cued (tone) conditioning and taste aversion protocols. These findings represent a rare instance of elevated learning and memorythat is localized and experimentally accessible to both in vitro and in vivo analyses, and is the only such case associated with fear and emotion learning. Mutations of this gene may be a contributing factor in human neuropsychiatric disorders characterized by increased excitatory activity inthe amygdala or in amygdala dependent circuits. This gene, and reagents developed from it, should provide a useful model system for the development of therapeutic interventions. (more...)

TCL1 is a protooncogene overexpressed in many mature B cell lymphomas. TCL1 is also expressed in precursor T cells and absent by the CD4+ CD8+ stage of thymocyte development. In B cells, TCL1 is first expressed in pro-B cells, and is completely absent in memory B cells and plasma cells. (more...)

Erythropoietin (EPO) is a cytokine that binds to its receptor (EPOR) and plays critical roles in hematopoiesis. Both Epo and EpoR are expressed in the nervous system during development and their expression levels are upregulated upon hypoxic injury. Exogenous EPO is neuroprotective and neurotrophic in CNS injury models. However, the endogenous role of EPO and the role of EPOR in neuroprotection and neurogenesis after stoke have not been identified.Stroke is the leading cause of adult disability due to the brains limited ability to repair after injury. Research tools to study the recovery of the brain after a stroke will aid in the development of future treatment options for stoke patients. (more...)

PTEN has been shown to be central in the regulation of cellular growth and survival. In many forms of cancer, mutations or loss of function in PTEN results in tumor formation - and as such PTEN is now appreciated as a central player in tumor suppression. Furthermore, PTEN has been shown to have a modulating role on cellular functions other than growth and survival, implicating this protein in diseases such as diabetes. Therefore, examination of PTEN in other models of disease may provide insight towards the understanding of mechanism and treatment of these diseases. (more...)

Researchers at UCLA have developed a C57Bl/6 based mouse model which lacks expression of a receptor for Type I Interferon (IFNAR). The animal lacks expression of the Type I receptor on all cells which has been shown to be the case genetically, biochemically and systemically in pathogen infection models. This is the first IFNAR KO mouse to be available on the C57Bl/6 background which is of most value to immunologists. (more...)

In the western world, prostate cancer remains the most common malignancy and the second-leading cause of cancer-related deaths in men. The PTEN gene (phosphotase and tensin homologue deleted on chromosome 10) has been well characterized as a tumor suppressor gene, and its malfunction/deletion has been linked to numerous cancer types, including prostate. Mutations or deletions of PTEN have been found in 30% of primary prostate cancers and 63% of malignant cases, ranking it as one of the most common determinants of prostate tumor progression. PTEN-controlled signaling pathways are therefore promising targets for therapeutic strategies, though significant problems in producing animal models have prevented serious gains toward this end. Lack of PTEN leads to embryonic lethality and heterozygote PTEN animals do not undergo normal prostate cancer development. A significant need remains for a workable PTEN-null prostate cancer animal model system. (more...)

B-raf is a member of the Raf family of intracellular signaling proteins, which also include Raf-1 and A-raf. Members of this family are involved in the all-important cellular signaling mechanism known as the Ras/Raf/MEK/ERK/MAPK signaling pathway. This ubiquitous pathway relays signals from outside the cell into the cells nucleus, regulating activities such as gene expression, differentiation, cell division, cell survival and cell death. B-raf regulates vital functions in the brain, testes, skin and bone marrow, and mutations in B-raf are found in malignant melanoma, certain types of thyroid and ovarian cancers, and sporadic types of colon cancers. Up to 6% of all human malignancies may harbor this mutation. Most recently, Sorafenib (Nexavar), which inhibits raf as well as a whole host of other signaling proteins, has been approved by the FDA for the treatment of advanced cancer of the kidney (clear-cell renal cell carcinoma). (more...)

Prostate cancer is the second leading cause of cancer deaths among men in the United States. Current treatments include surgery, radiation and anti-androgen therapy, although each option carries with it undesirable side-effects. Development of new pharmaceuticals with which to treat the disease relies on animal models that allow investigators to see easily the effects of a drug on the prostate. Current prostate cancer animal models require sacrificing the animal before the pharmacological effects of a drug can be observed. This makes following a drug at multiple time points tedious and expensive. While the use of cell lines simplifies assaying a drugs potency, it does not provide in situ information. Thus, the development of an animal model that would allow one to assay with ease the effectiveness of lead compounds could greatly accelerate development. (more...)

The MAGI-2 membrane protein (membrane-associated guanylate kinase inverted 3) acts as a scaffold to interact directly with and regulate the stability of both PTEN and IRSp53. In addition, members of the MAGI protein family are known to cluster receptors, signaling, and scaffolding proteins at specific membrane regions, including tight and synaptic junctions thereby playing important roles in regulating cell adhesion and signaling. (more...)

G protein-coupled receptor 4 (GPR4) has recently been identified as a novel proton-sensing receptor. GPR4 is expressed in vascular endothelial, smooth muscle, and several other cell types. UCLA researchers developed a GPR4 knockout mouse in order to better understand the in vivo role of GPR4 and investigate the hypothesis that GPR4 acts as a pH sensor in blood vessels. (more...)

The Myeloperoxidase (MPO) enzyme aids in the defensive properties of phagocytic cells of the human immune response. Prevalent in neutrophils and monocytes/macrophages, MPO generates a variety of oxidative processes which aid in the defensive mechanism of the host. Due to the fact that these cells are usually the primary responders to a diseased state, their defensive enzymes are often non-specific. In turn, the oxidative enzymes generated by MPO, may potentially play a role in the disease processes. (more...)

VIP and PHI are expressed at high levels in the neurons of the suprachiasmatic nucleus, the area of the nervous system responsible for most circadian behavior, but their function in the regulation of circadian rhythms is unknown. In order to study the role of these peptides on the circadian system in vivo, a new mouse model was developed in which both VIP and PHI genes were disrupted by homologous recombination. (more...)

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UC San Diego researchers have invented a device and method that provides objective evaluation of the reaction of mice to stimuli. Measurements can be made without human intervention or attendance. The device is engineered to be consistent, repeatable, and accurate, and can complete a measurement within 15 minutes. By rendering the measurements of reaction to stimuli objective, a high-throughput method for evaluating mouse genetic variation via this device can be made in a manner superior to behavioral analysis and other subjective methods. (more...)

There are several methods currently employed to quantify the behavioral reaction induced by an insult to an animal, generally to a paw. These include (1) observing the numbers of flinches of the affected paw; (2) measuring the time spent in different behavioral states (elevating, licking, biting, or shaking the affected limb); and (3) the use of weighted scores in which numerical weights are assigned to the different categories. The more complicated measures have not proven superior to the simple expedient of counting flinches. All of these procedures require continuous observation, severely limiting the number of animals that can be observed simultaneously. Since these observations are quantified subjectively, there is variability among individual observers. While several attempts have been made to automate the procedure, most of those attempts measure gross locomotor activity and not the isolated movement of the affected limb. (more...)

Caveolins are the principal protein components of caveolae, or “little caves,” which are 50 to 100 nm invaginations found in most cell types and represent appendages or subcompartments of plasma membranes. UC San Diego researchers have created a transgenic mouse for over-expressing caveolin-3 (M-caveolin) specifically within the cardiac myocyte. Caveolin-3 is one of the proteins that regulate signaling pathways and is thought to function in cardiac protection (ischemic preconditioning). Associated studies indicate that the novel caveolin-3 over-expressing mouse is resistant to myocardial ischemia/reperfusion injury, suggesting that caveolin-3 as a potential therapeutic for patients undergoing cardiac surgery or at high risk for myocardial infarction. (more...)

Animal models are useful not only for studying and understanding the biological and genetic factors that influence the development of different cancers, but also for developing treatments against those cancers. Unfortunately, although epithelial tissue derived cancers (e.g. breast, colon, lung, skin, and ovarian), are among the most common human cancers, very few models exist for studying the events leading to these tumor types or for testing new therapeutic treatments. Instead, many murine cancer models, incorporating mutations in tumor suppressor genes, typically develop soft tissue sarcomas or lymphomas. Therefore, a mouse model of epithelial carcinomas will facilitate the study and treatment of many common cancers not fully addressed by currently available models.Researchers at the University of California, San Francisco (UCSF), have developed a knockout mouse model of epithelial tissue derived carcinomas. This newly developed cancer model does not rely upon the breeding of multiple generations in order to develop chromosomal instability and is not dependent on homozygous loss of tumor suppressor genes. Instead, following exposure to ionizing radiation, tumor development occurs 2-5 months later and develops in multiple tissues including the ovary, lung, and liver. With their rapidity of tumor onset, wide spectrum of tumors, and decreased animal maintenance, these knockout mice represent a new model for studying ovarian as well as other epithelial derived cancer types and will be useful for the development of therapeutic treatments. (more...)

Background   Since the human genome has been sequenced, researchers are now focusing on defining the roles of each the ~30,000 genes in development and disease. As most human genes have homologs in the mouse genome, many researchers study the function of mouse genes to model the function of related human genes.       To study all aspects of gene function, one must be able to manipulate a single gene to generate many kinds of mutations: deletions, point mutations, insertion of exogenous DNA, etc. In mice, such site-directed mutagenesis is commonly achieved by homologous recombination of embryonic stem cells, which can be laborious, time-consuming, and inefficient. Thus, there is a need for technologies that allow for high-throughput, site-directed mutagenesis.   Description   UCSF investigators have designed vectors and methods for high-throughput genetic modification of an existing collection of gene trap insertions in mouse embryonic stem cells. The collection is publically available from the Sanger Gene Trap Consortium and currently consists of ~20,000 random insertions in ~4,500 genes in the mouse genome.   The vectors designed by UCSF investigators make use of recombinase activity to drive three recombination steps at the site of the gene trap insertion. The first step allows for reversion of the original gene trap insertion, resulting in the generation of an internal control. The final two steps allow for the insertion of new DNA of interest, resulting in the desired mutant or allele. Each recombination step can be selected easily using a visible genetic marker.   This system allows one to generate any number of mutations repeatedly into the same genomic site, with efficiencies as high as 80%. Additionally, because the insertions are controlled by the same endogenous promoter elements, position effects, which can create artifacts by misexpression or overexpression, are effectively eliminated. Using these vectors and methods, one could generate with relative ease a variety of site-directed mutations in any of the 4,500 genes for which there is an available enhancer trap insertion.     (more...)

University of California, Irvine researchers have developed a novel transgenic mouse model that contains the three major genes that contribute to the hallmark pathological features of Alzheimer's disease. These mice are exceedingly valuable for therapeutic investigations and for basic research aimed at understanding the behavioral, physiological, molecular/cell biological, and pharmacological processes leading to dementia in an animal model. Even though these mice contain three transgenes, the mice essentially breed as readily as a "single" transgenic line, greatly facilitating the establishment and maintenance of an animal colony. (more...)

This invention is an animal model and various cell-based models to determine nociception, pain transduction, and pain threshold. Researchers at the University of California, Irvine, discovered that overexpression of voltage-gated calcium channel alpha-2-delta-l subunit in neural tissue, and especially increased expression in spinal cord and dorsal root ganglia in transgenic mice, correlates in vivo with typical nerve injury-induced nociceptive responses to innocuous mechanical and thermal stimulation (tactile allodynia and thermal hyperalgesia). Significantly, such transgenic animals can be used to investigate nerve injury-induced neuropathic pain without inflicting nerve injury to the animals, which often interferes with test results. Similarly, neural cells obtained from such transgenic animals or neural cells transformed by overexpressing the alpha-2-delta-l subunit exhibit physiological parameters remarkably similar to those of neural tissue obtained from animals thought to have nerve injury-induced neuropathic pain. (more...)

Mgat5 is an enzyme that glycosylates T-cell receptors to modulate T-cell activation and autoimmune response. Mgat5 Null PL/J Mice were developed as part of a study performed by scientists at Mount Sinai Hospital and the University of California, Irvine to establish an understanding of the role of Mgat5 modified glycans in the regulation of autoimmunity. (more...)

BRCA2/RAD5 1 interaction is essential for DNA repair mechanisms and play significant role in tumor resistance to irradiation and chemotherapy treatments. Effective strategies to selectively interfere with BRCA2/RAD5 1 interaction in the context of treatment and chemoprevention of neoplastic diseases are described. (more...)

Sjogren's Syndrome is a grave autoimmune disease in which destruction of the exocrine glands leads to severe dryness of the eyes and mouth, with additional systemic complications. The pathogenesis of Sjogren's Syndrome is unclear, and treatment options are limited. Development of good animal models of the disease might lead to effective treatments for human patients, as well as a greater understanding of the cellular and molecular alterations that underlie disease progression. (more...)