MC1R (melanocortin 1 receptor) gene carrier and construction method thereof

Abstract

The invention discloses an MCIR (MelanoCortin 1 Receptor) gene carrier and a construction method thereof. The MCIR gene carrier comprises MC1R-neo and MC1R-GFP (Green Fluorescent Protein), wherein the nucleotide sequence of the MC1R-neo is SEQ ID NO:1, and the nucleotide sequence of the MC1R-GFP is SEQ ID NO:2. A CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) / Cas9 system is adopted to construct an MC1R gene knock-out vector, and only a sgRNA (Ribonucleic Acid) of which the length is about 20bp needs to be designed by aiming at a gene knock-out point to be connected with a universal Cas9 gene. Therefore, compared with traditional gene knock-out technologies including ZFNs (Zinc finger nuclease), TALENs (Transcription Activator-Like Effector Nucleases) and the like, the construction method disclosed by the invention adopting the CRISPR / Cas9 to construct the gene knock-out vector is simpler and more convenient and is more convenient to be further popularized and applied to subsequent experiments.

Description

technical field [0001] The invention belongs to the technical field of gene carriers, and in particular relates to an MClR gene carrier and a construction method thereof. Background technique [0002] As an important economic trait, the coat or plumage color of animals plays an important role in determining the purity and kinship of breeds. Studies have found that the different fur or feather colors of animals are caused by the different types and distributions of melanin. Melanocortin 1 receptor (MC1R), the main gene controlling the formation of animal melanin, consists of one exon, expressed in melanocytes, and interacts with the natural ligand a-MSH, through G The protein-coupled cAMP signaling pathway regulates a series of cascade reactions in melanocytes, and finally synthesizes their respective derivatives, pheomelanocytes and eumelanin, through dopa or dopachrome, thereby determining the coat color of animals. However, the molecular mechanism of MC1R affecting melan...

AI Technical Summary

Problems solved by technology

[0003] At present, in birds, the molecular mechanism of MC1R affecting melanin deposition is still unclear. In terms of its gene function, there are no reports on the knockout of MC1R gene and its effect on feather color traits.

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