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Knockout method for stem cell gene

A gene knockout and stem cell technology, applied in the field of molecular biology, can solve the problems of easy introduction of foreign genes, complex knockout methods, and inaccurate target gene function analysis, so as to achieve accurate target gene function analysis and good knockout effect , the effect of improving knockout efficiency

Pending Publication Date: 2020-04-28
CHANGZHOU NO 2 PEOPLES HOSPITAL
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Problems solved by technology

[0006] Aiming at the deficiencies in the prior art, the present invention provides a stem cell gene knockout method, which solves the problem that the existing stem cell gene knockout method is relatively complicated in the actual operation process, and at the same time, it is easy to introduce stem cell gene knockout. Exogenous genes make the follow-up personnel's analysis of the function of the target gene inaccurate

Method used

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Embodiment Construction

[0021] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some of the embodiments of the present invention, but not all of them. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

[0022] The embodiment of the present invention provides a technical solution: a stem cell gene knockout method, through which the stem cell gene knockout method adopted in the present invention can rapidly knock out stem cell genes without introducing foreign genes, and knockout The effect is good, so that the follow-up personnel can analyze the function of the target gene more accurately, improve the efficiency of gene knockout, and the operation is simple and repeatable, which meets the actual needs of use. Specifically, it include...

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Abstract

The invention discloses a knockout method for a stem cell gene, and relates to the technical field of molecular biology. The method specifically includes the following steps: S1, establishing a gene sequence, querying a code sequence (CDS) of a target gene in a database according to the gene sequence, obtaining a complete CDS of the gene by using a single-stranded CDNA as a template, and performing comparison to obtain a complete target gene exon sequence; S2, designing the target gene according to the S1, designing homologous recombination double exchange homologous arms, and constructing target gene knockout plasmid on the basis of knocking out template plasmid; and S3, recombining the target gene and DNA molecules homologous with a specific fragment of a target gene in cells onto a vector with a marker gene. According to the knockout method for the stem cell gene, the stem cell gene can be quickly knocked out on the basis of not introducing exogenous genes by using the knockout method for the stem cell gene provided by the invention, so that a worker can analyze the target gene function more accurately in the later stage, and the gene knockout efficiency is improved.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a stem cell gene knockout method. Background technique [0002] Stem cells are a type of cells with unlimited or immortal self-renewal ability, capable of producing at least one type of highly differentiated offspring cells, the function of stem cell population is to control and maintain cell regeneration, generally speaking, in stem cells Between them and their terminally differentiated progeny cells there are intermediate populations of progenitor cells known as "committed progenitors", which have limited expansion capacity and restricted differentiation potential, and the function of these cell populations is to increase stem cells each time The number of differentiated cells produced after division. Stem cells have the ability to self-renew, but the division of stem cells is actually relatively asymmetric. Although the structural significance of stem cells at the le...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85C12N15/65
CPCC12N15/65C12N15/85
Inventor 白阳
Owner CHANGZHOU NO 2 PEOPLES HOSPITAL
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