Demethylation-based vector for promoting self-renewal and proliferation of germline stem cells and application thereof

Abstract

The invention discloses a demethylation-based vector for promoting self-renewal and proliferation of germline stem cells and application thereof. The vector is coated in slow virus, and carries mTet1 gene. The vector is a slow virus vector, and the nucleotide sequence of the mTet1 gene is disclosed as SEQ.ID.NO.1. After the vector disclosed by the invention is exprssed through the slow virus, the mTet1 gene is integrated to the genome of the milk goat male germline stem cells. When the stable positive cells are cultured, the cells grow in a swirl cluster mode and have favorable cytomorphology; the stream-mode analysis on the cell division cycle detects that the quantity of S-stage cells is increased, which indicates that the proliferation capacity is enhanced; and the transcription level detects that the expression of the male germline stem cell related gene and self-renewal related gene is increased, and the expression of the differentiation related gene is decreased.

Description

technical field [0001] The invention belongs to the field of transgenic technology, and relates to a carrier for promoting the self-renewal and proliferation of reproductive stem cells based on demethylation and its application, especially acting on the male reproductive stem cells of dairy goats. Background technique [0002] Dairy goats are important economic animals in my country, and diseases such as macrofetal syndrome, high lethality and stunting in production have seriously hindered the maintenance and optimization of excellent animal breeds. These disorders are largely associated with abnormalities in spermatogenesis and embryonic development. Therefore, improving the quality of sperm and embryos, reducing the occurrence of diseases, and promoting the inheritance and breeding of fine varieties are technical problems that need to be solved urgently in the field of genetic breeding of large livestock. The establishment of DNA methylation during mammalian spermatogenes...

AI Technical Summary

Problems solved by technology

At present, Tet1-deficient ESCs and nude mice have been obtained. Studies have found that Tet1-deficient ESCs still maintain a totipotent state. Tet1-deficient nude mice are healthy and have reproductive capacity, but have defects in birth weight ratio, and neutrophils Compared with intraspecific hybrids, the litter size is relatively small, indicating that Tet1 plays a role in embryonic development and spermatogenesis (Dawlaty et al., 2011)

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