Method for realizing animal sex control by editing X chromosome multi-copy genes

An X chromosome and multi-copy technology, applied in the field of genetic engineering, can solve the problems of gender control effect to be further improved and high off-target rate, achieve good sex control effect, low off-target rate, and increase birth rate

Active Publication Date: 2020-08-18
SOUTH CHINA AGRI UNIV
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Problems solved by technology

However, the off-target rate of this method is high, an

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  • Method for realizing animal sex control by editing X chromosome multi-copy genes
  • Method for realizing animal sex control by editing X chromosome multi-copy genes
  • Method for realizing animal sex control by editing X chromosome multi-copy genes

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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 realizing animal sex control by performing gene editing on X chromosome multi-copy genes. The method comprises the following steps: screening and selecting one ormore multi-copy genes on an X chromosome of a mammal as target sites, and designing and synthesizing corresponding sgRNA; then, constructing a CRISPR/Cas9 expression system of a targeted X chromosomemulti-copy gene to cut the X chromosome in sperms or fertilized eggs of mammals; according to the method, the whole X chromosome can be inactivated, X sperms can be killed, the birth rate of male mammals can be increased, or XY type embryos can be killed, and the birth rate of female mammals can be increased. According to the method provided by the invention, the X chromosome cutting efficiency ishigh, the off-target rate is low, and a good sex control effect is achieved.

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 ...

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Application Information

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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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