Method for increasing pork yield by CRISPR/Cas9

A technology for meat production and floret pigs, applied in the field of genetic engineering, can solve the problems of germplasm genetic characteristics without high lean meat rate, slow increase in lean meat rate, and long measurement cycle, so as to avoid biosafety risks and improve lean meat. Meat rate, increased production and yield effects

Inactive Publication Date: 2018-12-25
SUN YAT SEN UNIV
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Problems solved by technology

[0003] However, the traditional breeding of lean meat percentage requires a large number of growth and slaughter performance measurements, the measurement cycle is long, the measurement cost is high, and the lean meat percentage increases slowly
This is mainly due to the fact that traditional breeding mainly considers the selection of comprehensive traits, and does not select the trait of lean meat percentage as a single trait; the data of lean meat percentage traits of pigs can only be obtained through slaughtering measurements, and usually the lean meat percentage of live pigs can only be obtained. Estimated by backfat thickness and eye muscle area data; Chinese local pig breeds do not have germplasm genetic characteristics of high lean meat rate and other reasons

Method used

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  • Method for increasing pork yield by CRISPR/Cas9
  • Method for increasing pork yield by CRISPR/Cas9
  • Method for increasing pork yield by CRISPR/Cas9

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Experimental program
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Effect test

Embodiment

[0028] 1. IGF2 gene target site selection

[0029] According to the IGF2 gene sequence and the basic design principles of sgRNA (the sequence conforms to 5'-GN19NGG-3'), and with the help of software analysis (http: / / crispr.mit.edu / ), a sgRNA targeting porcine IGF2 gene was designed. See Table 1 for the specific sequence.

[0030] Table 1 Sequence information of gRNA targeting IGF2 gene

[0031]

[0032] Note: PAM (protospacer-adjacent motif) refers to the sgRNA binding motif

[0033] 2. Construction of porcine IGF2 gene sgRNA / Cas9 targeting vector

[0034] The pX330 vector can simultaneously express sgRNA and codon-optimized human Cas9 protein.

[0035] According to the two sgRNAs designed above, the corresponding positive and negative single strands of oligonucleotides were synthesized, and corresponding BbsI sticky ends were added to each single strand. Digest 1 μg of the pX330 vector with BbsI, and after electrophoresis of the linearized plasmid, recover the digeste...

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Abstract

The invention provides a method for increasing pork yield by CRISPR / Cas9. The method includes:adopting CRISPR / Cas9 technique for constructing a targeting vector containing swine IGF2 gene targeting sgRNA, transfecting swine cells to obtain cells containing editing type IGF2 genes, and performing somatic cell nuclear transfer and embryo transfer, wherein a nucleotide sequence of the sgRNA for constructing the targeting sector is as shown in SEQ ID NO.1. The method has advantages that an IGF2 gene expression inhibiting effect of ZBED6 can be relieved, IGF2 expression is promoted, and the lean meat rate and the meat yield of pigs are increased.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and in particular relates to a method for improving pig meat production by using CRISPR / Cas9. Background technique [0002] In the field of domestic pig genetics and breeding in China, the traditional lean meat rate selection method is usually adopted. The implementation steps of this technology are individual lean meat rate measurement, BLUP estimated breeding value, and comprehensive ranking to select individuals. [0003] However, the traditional breeding for lean meat percentage requires a large number of growth and slaughter performance measurements, the measurement cycle is long, the measurement cost is high, and the lean meat percentage increases slowly. This is mainly due to the fact that traditional breeding mainly considers the selection of comprehensive traits, and does not select the trait of lean meat percentage as a single trait; the data of lean meat percentage traits of pigs can...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85C12N15/90C12N15/66C12N15/113A01K67/027
CPCA01K67/0276A01K2217/075A01K2227/108A01K2267/02C07K14/65C12N15/113C12N15/66C12N15/8509C12N15/907C12N2310/10C12N2800/107C12N2810/10C12N2310/20
Inventor 何祖勇陈瑶生刘小红刘小凤
Owner SUN YAT SEN UNIV
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