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Method for editing x-chromosome multi-copy genes to realize animal sex control

An X-chromosome, multi-copy technology, applied in the field of genetic engineering, can solve the problems of high off-target rate and the effect of gender control needs to be further improved.

Active Publication Date: 2022-05-13
SOUTH CHINA AGRI UNIV
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Problems solved by technology

However, the off-target rate of this method is high, and the gender control effect needs to be further improved

Method used

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  • Method for editing x-chromosome multi-copy genes to realize animal sex control
  • Method for editing x-chromosome multi-copy genes to realize animal sex control
  • Method for editing x-chromosome multi-copy genes to realize animal sex control

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Embodiment 1

[0039] 1. Selection of mouse multi-copy gene target sites and design of corresponding sgRNA

[0040] The X staining of the mouse was deeply sequenced, and the repeated sequences with more than 50 copies were selected for use. Then, the CRISPR / Cas9 target sequence online design software was used to analyze the candidate sequence, and compared with the whole mouse genome sequence, the X-B and X-D sites with the lowest off-target efficiency were selected, and the corresponding sgRNA X-B and sgRNA X-D sequences were designed. The information is shown in Table 1-2 below.

[0041] Table 1 Multi-copy target loci on mouse X chromosome

[0042] target site name Sequence (5'-3' underline marked as PAM region) copy number X-B GCTTGGTTAGGGTGAGTGCT TGG (SEQ ID NO:1)

72 X-D TAAGTGCTGTGTGCTGCTAC TGG (SEQ ID NO:2)

99

[0043] Table 2 sgRNA sequence

[0044] sgRNA name target site Sequence (5'-3') sgRNA X-B X-B GCTTGGTTAGGGT...

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Abstract

The invention discloses a method for gene editing of X chromosome multi-copy genes to realize animal sex control. By screening and selecting one or more multi-copy genes on the X chromosome of mammals as target sites, designing and synthesizing corresponding sgRNAs, and then By constructing a CRISPR / Cas9 expression system targeting multiple copies of the X chromosome to cut the X chromosome in mammalian sperm or fertilized eggs, the entire X chromosome can be inactivated, the X sperm can be killed, and the birth rate of male mammals can be improved. , or lethal XY embryos, increase the birth rate of female mammals. The method provided by the invention has high cutting efficiency of X chromosome, low off-target rate, and good sex control effect.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a method for realizing sex control of mammals through gene editing of X chromosome multi-copy genes. Background technique [0002] In order to meet different production needs, people are eager to obtain a large number of livestock and poultry of the required sex through sex control technology, and reduce the waste of raw materials. The gene editing technology is a technology for editing the genome of an organism, which can realize the knockout, insertion and replacement of a certain gene, which provides a new direction for the research of sex control technology. Using gene editing technology, a knockout line of sex-related genes can be established to study its role in sex differentiation and gonad development, and then the expression of sex-related genes can be enhanced or inhibited by inserting foreign genes to achieve the purpose of sex ratio shift . However, the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/90C12N15/85A01K67/027
CPCC12N15/907C12N15/8509A01K67/0276A01K2267/03A01K2227/105A01K2217/075
Inventor 吴珍芳李紫聪谢绍怡蔡更元郑恩琴杨化强杨杰洪林君顾婷徐铮黄思秀
Owner SOUTH CHINA AGRI UNIV
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