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SgRNA targeting sequence specifically targeting mouse G6pc gene and application thereof

A kind of specific, mouse technology, applied in the fields of medical genetics and molecular biology, can solve the problem of no sgRNA guide sequence, knockout, etc., to achieve high-efficiency gene editing, increase the number, and improve the efficiency of cell transfection.

Active Publication Date: 2020-03-13
GUILIN MEDICAL UNIVERSITY
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  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

However, there is currently no sgRNA guide sequence for the mouse G6pc gene and a method for knocking out the gene

Method used

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  • SgRNA targeting sequence specifically targeting mouse G6pc gene and application thereof
  • SgRNA targeting sequence specifically targeting mouse G6pc gene and application thereof
  • SgRNA targeting sequence specifically targeting mouse G6pc gene and application thereof

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Embodiment Construction

[0079] The principles and features of the present invention will be described below in conjunction with specific drawings, and the examples given are only used to explain the present invention, not to limit the scope of the present invention.

[0080] 1. Screening of human G6PC gene pathogenic mutation sites, confirming that Arg at position 83 and Arg at position 170 of the human G6PC gene are potential mutation sites

[0081] Glycogen storage disease type I is caused by mutations in the G6PC gene, and 14 pathogenic mutations were screened using the OMIM online database (as shown in Table 1). Due to the limited mutation sites included in OMIM, the present invention also screened the function inactivation mutation of the human G6PC gene using the ExAC database, and screened 11 mutation sites in total (as shown in Table 2 and figure 1 shown). Then use the ClinVar database to retrieve and screen the 25 mutation sites to determine their mutational pathogenicity (such as figure ...

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Abstract

The invention discloses a sgRNA targeting sequence specifically targeting the mouse G6pc gene and application thereof, and belongs to the technical field of medical genetics and molecular biology. Thenucleotide sequence corresponding to the sgRNA contains the sequence as shown in SEQ ID NO. 2. The invention also discloses a method for editing the mouse G6pc gene by using the above sgRNA targetingsequence specifically targeting the mouse G6pc gene. The sgRNA targeting sequence of the invention can mediate the Cas9 protein and efficiently cut target DNA, and further can be used for editing themouse G6pc gene to affect the function of mouse G6pc gene encoding protein. The sgRNA targeting sequence can efficiently realize targeting by a CRISPR / Cas9 system with an efficiency of 100%.

Description

technical field [0001] The invention relates to a sgRNA guide sequence specifically targeting mouse G6pc gene and application thereof, belonging to the technical fields of medical genetics and molecular biology. Background technique [0002] Clustered regularly interspaced short palindromic repeats (Clustered regularly interspaced shortpalindromic repeats, associated RNA guided endonuclease Cas, CRISPR / Cas) is an acquired immune defense system formed by bacteria and archaea in the long-term evolution process against foreign genetic material. Among them, the type II CRISPR / Cas9 system has a relatively simple structure and has been widely used for gene editing in various species through genetic engineering. The system consists of two components, Cas9 protein and sgRNA. The sgRNA binds to the complementary sequence of about 20 nt in genomic DNA through complementary base pairing, and mediates the HNH active site and RuvC active site of Cas9 protein to cut DNA. double strand, r...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/85C12N15/90C12N9/22A01K67/027
CPCA01K67/0275A01K2217/07A01K2227/105C12N9/22C12N15/113C12N15/85C12N15/907
Inventor 付灿于鸿浩岳鹏鹏李勇农月娟
Owner GUILIN MEDICAL UNIVERSITY
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