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Light-inducible Cre Transgenic Mouse Models

光诱导 Cre 转基因小鼠模型

基本信息

批准号：
8767916
负责人：
Richard R Behringer
金额：
$ 24万
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 2016-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8767916
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 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 human disease in vivo loss of function mouse model novel phyB phytochrome phycobilin postnatal public health relevance 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用于细胞类型特异性和/或时间模式中的基因表达激活。这些方法通常依赖于确定的细胞类型特异性或普遍存在的基因调控序列来指导Cre或CreER（他莫昔芬诱导型）表达。因此，缺乏相关基因调控序列或受他莫昔芬影响的细胞类型或组织区域难以进行遗传修饰，阻碍了对这些细胞类型或组织中基因功能的研究。经典的胚胎学操作已被用于在缺乏特定基因调控序列的情况下修饰发育中的组织。例如，浸泡在分泌蛋白质中的珠子可以物理植入特定阶段发育胚胎的精确区域，以改变组织分化。类似地，稳定表达分泌的蛋白质编码基因可以沉淀，然后移植到胚胎区域以改变宿主组织分化。然而，这些胚胎学方法是有限的，因为它们仅限于检查编码分泌蛋白质的基因或不由基因编码的分子（例如视黄酸），并且因为移植的珠粒或细胞团块必然破坏发育中的胚胎组织的天然结构。此外，珠粒和细胞团块移植只能在有限的情况下进行。我们提出了一种遗传方法，该方法独立于定义的细胞类型特异性调控序列来表达任何基因，该基因还保持组织结构以绕过标准Cre/lox方法的先前困难。这将为小鼠的条件遗传获得和功能丧失方法开辟新的可能性。我们的方法也适用于其他模式生物。该提议的主要目的是产生表达光诱导Cre系统的转基因小鼠，以与任何当前Cre依赖性获得和丧失功能等位基因联合收割机组合，以研究各种生物过程和人类疾病模型。