Inducible CRISPRon or CRISPRi mouse embryo stem cells and application thereof

A mouse embryo and stem cell technology, applied in embryonic cells, cells modified by introducing foreign genetic material, applications, etc., can solve problems such as genome changes, and achieve stable expression, high homologous recombination efficiency, and high efficiency.
CN107142247AActive Publication Date: 2017-09-08TIANJIN MEDICAL UNIV
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
TIANJIN MEDICAL UNIV
Publication Date
2017-09-08

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Abstract

The invention provides inducible CRISPRon or CRISPRi mouse embryo stem cells and a preparation method thereof. The mouse embryo stem cells are A2Lox.Cre cells and can reversibly express dCas9-VPR fusion protein or dCas9-KRAB fusion protein. The invention further provides a method for regulating and controlling gene expression and a kit. According to the inducible CRISPRon or CRISPRi mouse embryo stem cells provided by the invention, the construction method is simple and convenient, and the stable positive rate is up to 90 percent or higher; the genomic sequence is not edited, and gene expression is directly activated or inhibited; expression of the fusion protein has inducibility and reversibility; controllability and diversity on targeted regulation and control on the gene are achieved.
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Description

technical field

[0001] The invention relates to the field of gene expression regulation, in particular to an inducible CRISPRon and CRISPRi mouse embryonic stem cell and its application. Background technique

[0002] RNA interference (RNAi) or gene overexpression (overexpression, OE) is a common method for gene function research. However, RNAi is a post-transcriptional regulation and is prone to off-target phenomena; overexpression (OE) often uses the promoter of the cloning vector to promote the expression of foreign genes, and the activity of the promoter is unstable and uncontrollable in different cell types.

[0003] The combination of regularly clustered interspaced short palindromic repeats CRISPR and endonuclease Cas9 can target and cut the DNA genetic material of invaders under the guidance of guide RNA (sgRNA). In 2012, researchers took advantage of this feature to make the CRISPR system into a powerful genome editing tool (Jinek, M. et al., 2012; Cong, L. et al., ...

Claims

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