Preparing method for MC3R gene knockout pigs

Abstract

The invention discloses a preparing method for MC3R gene knockout pigs. The preparing method for MC3R gene knockout pigs includes the steps that a CRISPR / Cas9 system is adopted, the CRISPR / Cas9 system comprises an encoding gene of gRNA1 and an encoding gene of gRNA2, and the target sequence of a MC3R gene identified by the gRNA1 is a sequence 1 in a sequence table; the target sequence of the MC3R gene identified by the gRNA2 is a sequence 2 in a sequence table. An experiment shows that the MC3R gene knockout efficiency is 29.16% with the preparing method for MC3R gene knockout pigs; a target gene can be rapidly and efficiently knocked out in a large-fragment mode, exogenous gene fragments are not left, and the preparing method can be used for researching the action mode and mechanism of the MC3R gene, and can be further used for animal breeding.

Description

technical field [0001] The invention relates to a preparation method of MC3R gene knockout pigs in the field of biotechnology. Background technique [0002] Melanocortin receptors (MCRs) are receptors located on the surface of mammalian cells, belonging to the G-protein coupled receptor family, and five members have been discovered so far. Under the stimulation of Melanocortin (MC), MCRs increase the concentration of the intracellular second messenger cAMP by binding to the activated Gs protein, and start the intracellular signaling pathway. The physiological role of each member of MCRs is very different. MC1R is expressed by skin melanocytes and plays a key role in determining skin and hair pigmentation. MC2R is expressed in the adrenal cortex and regulates adrenal steroid synthesis and cell proliferation. MC5R is widely distributed and may be related to exocrine function. MC3R and MC4R are believed to be involved in the regulation of energy balance in the body. In rece...

AI Technical Summary

Problems solved by technology

The construction of this targeting carrier is cumbersome, the cycle is long, the cost is high, and the efficiency is extremely low

And the deletion of mark gene is a troublesome problem, even after mark-free, the gene fragment of loxp or flt will be left

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