Specifically-targeted swine IGFBP3 gene sgRNA targeting sequence and application

A kind of specific and sequence technology, applied in sgRNA guide sequence and application field, can solve the problems of mouse weight gain, expression level increase, content increase, etc., and achieve high safety and eliminate expression effect
CN107099533AInactive Publication Date: 2017-08-29NORTHEAST AGRICULTURAL UNIVERSITY
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
NORTHEAST AGRICULTURAL UNIVERSITY
Publication Date
2017-08-29
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention relates to a sgRNA targeting sequence and application, in particular to a specifically-targeted swine IGFBP3 gene sgRNA targeting sequence and application. The sgRNA targeting sequence is CGTCTCGCGCTTGGACTCAG. By the sgRNA targeting sequence, an IGFBP3 gene can be knocked out or edited through a CTISPR / Cas9 system so as to eliminate IGFBP3 expression to lay the foundation for preparation of IGFBP3 transgenic swine. The specifically-targeted swine IGFBP3 gene sgRNA targeting sequence is applied to the field of gene engineering.
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Description

technical field

[0001] The invention relates to a sgRNA guide sequence and its application. Background technique

[0002] The CRISPR / cas9 gene editing system is modified based on the class II CRISPR / Cas system. Cas9 protein is the only Cas protein required to mediate silencing of foreign DNA. In 2012, Jinek confirmed through in vitro experiments that the cutting of the protospacer sequence by the Cas9 protein requires not only a mature crRNA complementary to the protospacer sequence, but also tracrRNA; by sequencing the cleaved fragments, it was found that tracrRNA:crRNA guides the cleavage site of the Cas9 protein The point is site-specific, whether it is a circular plasmid or a linear DNA fragment, the cutting site is at the 3bp upstream of the PAM sequence; the Cas9 protein contains HNH and RuvC endonuclease domains, no matter which domain is mutated , are unable to cut effectively, but only produce nicks on double-stranded DNA. Experiments have confirmed that the HNH d...

Claims

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