Construction method for conditional overexpression Yap1 gene mice model at H11 locus

A construction method and mouse model technology, applied in the field of genetic engineering, can solve problems such as the lack of a clear understanding of basic issues in developmental biology, and achieve the effects of increasing the probability of homologous recombination repair, improving reliability, and improving accuracy

Inactive Publication Date: 2019-08-06
成都药康生物科技有限公司
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Problems solved by technology

However, despite intensive research on the rationale behind organs of "preset" size, this fundamental question in developmental biology has not yet been clearly understood

Method used

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  • Construction method for conditional overexpression Yap1 gene mice model at H11 locus
  • Construction method for conditional overexpression Yap1 gene mice model at H11 locus
  • Construction method for conditional overexpression Yap1 gene mice model at H11 locus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0022] Example: Construction of conditionally overexpressed mouse Yap1 animal model at the H11 site

[0023] Using CRISPR Cas9 gene editing technology to knock in the Yap1 gene sequence at the H11 site of the mouse model, and at the same time achieve conditional overexpression of Yap1, so as to establish a conditional overexpression of Yap1 mouse model. C57BL / 6J mice are a well-developed mouse strain at present. Using C57BL / 6J mice as background mice, a conditional Yap1 mouse model was successfully obtained. The acquisition of the model includes the following steps:

[0024] (1) Determine the H11 site

[0025] According to the mouse chromosome 11 sequence, the gene sequence of the H11 site was selected, and the selected gene sequence was shown in SEQ.ID.NO.1.

[0026] (2) sgRNA screening of conditionally overexpressed mouse Yap1 at the H11 site

[0027] The sgRNA targeting the mouse sequence was designed at the H11 site. Design and synthesize the recognition sgRNA sequence...

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Abstract

The invention discloses a construction method for a conditional overexpression Yap1 gene mice model at an H11 locus. The construction method comprises the steps: firstly, obtaining an inducible Yap1 overexpression transgenic vector; then, obtaining transgenic F0-generation mice; next, enabling the F0-generation mice to mate with wild-type mice, carrying out passage and line establishment, and thusobtaining F1-generation mice; and carrying out expanding propagation of positive F1-generation mice, then mutually mating, and thus obtaining the conditional overexpression mice Yap1 animal model. The transgenic mice model constructed by the method can be used for studying the function of Yap1 in different tissues and different developmental stages and the action mechanism of Hippo signaling pathways, can also be used for studying the function of the Hippo signaling pathway in animal development, biotumorigenesis, regenerative medicine and other disciplines, and has important guiding significance on biological signal transduction studies.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a method for constructing a mouse model for conditionally overexpressing Yap1 gene at the H11 site. Background technique [0002] Complex biological processes often require in vivo analysis, and the study of human in vivo biology is severely constrained by ethics and technology, so animal models are increasingly required for in vivo studies of human cells, tissues, and organs. [0003] After two decades of research, the Hippo signaling pathway has been identified in a variety of model organisms as a key factor regulating organ size and tissue homeostasis during development. Through repression of YAP and TAZ transcriptional coactivators, the Hippo pathway regulates cell proliferation, apoptosis, and stemness in response to a wide range of extracellular and intracellular signals, including cell-cell contact, cell polarity, mechanosignaling, G proteins Ligan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85A01K67/027
CPCA01K67/0275A01K2217/05A01K2227/105A01K2267/03C07K14/47C12N15/85
Inventor 史培良高翔王宏宇
Owner 成都药康生物科技有限公司
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