Light-inducible Cre Transgenic Mouse Models

光诱导 Cre 转基因小鼠模型

基本信息

批准号：
8908066
负责人：
Richard R Behringer
金额：
$ 19.43万
依托单位：
UNIVERSITY OF TX MD ANDERSON CAN CTR
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-08-10 至 2017-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8908066
关键词：
Adult Alleles Animal Model Arabidopsis Architecture Binding Biological Models Biological Process Biomedical Research Bypass C-terminal Cells Chemicals Chimeric Proteins Communities Development Disease Embryo Embryonic Development Embryonic Lethal Mutation Engineering Family member Gene Expression Gene Silencing Gene Targeting Generations Genes Genetic Genetically Engineered Mouse Health Homeostasis Implant In Vitro Lead Light Methods Modeling Mus Mutate Organ Organ Culture Techniques Pattern Phytochrome Proteins Regulator Genes Reporter Research Staging System Tamoxifen Testing Tissue Differentiation Tissues Transcription Coactivator Transgenic Mice Transplantation Tretinoin cell type embryo tissue gene function genetic approach human disease in vivo loss of function mouse model novel phyB phytochrome phycobilin postnatal reproductive response success tissue/cell culture tool

项目摘要

DESCRIPTION (provided by applicant): Cre/lox conditional genetic approaches in the mouse are used for cell type-specific and/or temporal gene inactivation to bypass early embryonic lethal mutations or for activation of gene expression in cell type-specific and/or temporal patterns. These approaches generally rely on defined cell type-specific or ubiquitous gene regulatory sequences to direct Cre or CreER (tamoxifen-inducible) expression. Cell types or tissue regions that lack associated gene regulatory sequences or are influenced by tamoxifen are therefore difficult to genetically modify, hindering studies of gene function in these cell typs or tissues. Classical embryological manipulations have been used to modify developing tissues in the absence of specific gene regulatory sequences. For example, beads soaked in secreted proteins can be physically implanted into a precise region of a developing embryo of a specific stage to alter tissue differentiation. Similarly, tissue culture cells stably expressing a secreted protein-encoding gene can be pelleted and then transplanted into an embryonic region to alter host tissue differentiation. However, these embryological approaches are limited because they are restricted to examining genes encoding secreted proteins or molecules not encoded by genes (e.g. retinoic acid) and because the transplanted bead or cell pellet necessarily disrupts the natural architecture of the developing embryonic tissue. In addition, bead and cell pellet transplants can only be performed in a limited set of situations. We propose a genetic approach that is independent of defined cell type-specific regulatory sequences to express any gene that also maintains tissue architecture to bypass previous difficulties with standard Cre/lox approaches. This should open up novel possibilities for conditional genetic gain- and loss-of-function approaches in the mouse. Our approach should also be applicable to other model organisms. The primary objective of this proposal is to generate transgenic mice that express a light-inducible Cre system to combine with any of the current Cre-dependent gain- and loss-of-function alleles to study a wide variety of biological processes and model human diseases.

描述（由申请人提供）：小鼠中的CRE/LOX条件遗传方法用于细胞类型特异性和/或时间基因失活，以绕过早期胚胎致死突变或在细胞类型特异性和/或时间模式中激活基因表达。这些方法通常依赖于定义的细胞类型特异性或普遍存在的基因调节序列来指导Cre或CRER（他莫昔芬诱导）表达。因此，缺乏相关基因调节序列或受他莫昔芬影响的细胞类型或组织区域很难基因修饰，这阻碍了这些细胞类型或组织中基因功能的研究。在没有特定基因调节序列的情况下，经典的胚胎操纵已用于修饰发展组织。例如，浸泡在分泌的蛋白质中的珠子可以物理地植入特定阶段的发育中的胚胎的精确区域，以改变组织分化。同样，组织培养细胞稳定表达分泌的蛋白质编码的基因可以颗粒，然后移植到胚胎区域以改变宿主组织分化。但是，这些胚胎学方法受到限制，因为它们仅限于检查编码未由基因（例如视黄酸）编码的分泌蛋白质或分子的基因，并且因为移植的珠子或细胞颗粒一定会破坏发育中胚胎组织的自然结构。另外，只能在有限的情况下进行珠子和细胞颗粒移植。我们提出了一种独立于定义的细胞类型特异性调节序列的遗传方法，以表达任何还保持组织结构以绕过以前的CRE/LOX方法绕过以前的困难的基因。这应该为小鼠中有条件的遗传增益和功能丧失方法打开新的可能性。我们的方法也应适用于其他模型生物。该提案的主要目的是生成转基因小鼠，该小鼠表达光诱导的CRE系统，以与当前CRE依赖性的功能丧失和功能丧失等位基因相结合，以研究各种生物学过程和模型人类疾病。