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Application of SCR7 in editing receptor genes in base editing system

A base editing and genome technology, applied in genetic engineering, vectors, viruses/phages, etc., can solve the problems of low efficiency and limitations of HDR

Active Publication Date: 2019-04-23
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For base-accurate substitution, the application of HDR to achieve base-accurate substitution is greatly limited due to the low efficiency of HDR and the need for DNA templates

Method used

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  • Application of SCR7 in editing receptor genes in base editing system
  • Application of SCR7 in editing receptor genes in base editing system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1. Effects of different concentrations of SCR7 treatment on the SpCas9n(D10A)&PmCDA1&UGI base editing system

[0048] 1. Construction of gene editing vector

[0049] The inventors of the present invention constructed the vector SpCas9n&PmCDA1&UGI.

[0050] The nucleotide sequence of the vector SpCas9n&PmCDA1&UGI (circular) is shown in sequence 1 in the sequence listing. In sequence 1, from 5' to 3', the 131st to 467th position is the nucleotide sequence of the OsU3 promoter, the 647th to 723rd position and the 820th to 896th position are all the nucleotide sequences of pre-tRNA, the first The 744th to 819th and 917th to 992nd are the nucleotide sequence of the sgRNA backbone, the 724th to 743rd is the nucleotide sequence of CS651, the 897th to 916th is the nucleotide sequence of CS652, and the 993rd to 1283rd The position is the nucleotide sequence of the OsU3 terminator, the 1290th to 3003rd is the nucleotide sequence of the OsUbq3 promoter, the 3010th to 727...

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Abstract

The invention discloses application of SCR7 in editing receptor genes in a base editing system. Through environments, it is proved that SCR7 can improve the editing efficiency of the base editing system and expand the mutation window range of the base editing system, and then a mutant containing more mutation sites is obtained. The application has an important application value.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of SCR7 in base editing system editing receptor genome. Background technique [0002] CRISPR-Cas9 technology has become a powerful genome editing method and has been widely used in many tissues and cells. The CRISPR / Cas9protein-RNA complex is positioned on the target by the guide RNA (guide RNA), cuts and generates a DNA double-strand break (dsDNA break, DSB), and then the organism will instinctively initiate a DNA repair mechanism to repair the DSB. There are generally two repair mechanisms, one is non-homologous end joining (NHEJ), and the other is homologous recombination (homology-directed repair, HDR). Usually NHEJ accounts for the majority, so the random indels (insertions or deletions) generated by the repair are much higher than the precise repair. For precise base substitution, the application of HDR to achieve precise base substitution is great...

Claims

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Application Information

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IPC IPC(8): C12N15/82
CPCC12N15/8213C12N2810/10
Inventor 杨进孝武莹宋伟袁爽王飞鹏
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES