动物实验,CRISPR-Cas9,基因敲除 z-bo 2020-09-28 708

　　Recently, researchers from Harvard University in the United States published the latest research results in the international academic journal "Development Cells", and used a vector system based on CRISPR-Cas9 technology to achieve zebrafish tissue-specific gene knockout. It is undoubtedly good news for scientists who have achieved that the main research tool is fish. Zebrafish has become an important model organism in life science research due to its easy reproduction, short reproduction cycle, large number of eggs, in vitro fertilization of embryos, in vitro development and transparent embryonic bodies. The platform. Zebrafish can be used to study the basic problems of life sciences and to clarify the molecular mechanisms of embryo and tissue organ development. You can establish various human disease and tumor models, and establish a research platform for drug screening and treatment. Can establish toxicology and aquatic reproduction. Model, research and solve key issues in environmental and agricultural sciences. As a gene editing tool,

　　CRISPR/Cas9 technology is widely used in a variety of platforms and systems, significantly promoting the knockout of target genes in vivo and in vitro. In this study, the researchers developed a vector system based on CRISPR-Cas9 technology to achieve tissue-specific gene knockout in zebrafish. They used CRISPR/Cas9 technology in zebrafish, and they only need to inject guide RNA (gRNA) and Cas9 mRNA at the single-cell embryo stage to quickly obtain gene knockout zebrafish. In order to prove the applicability of the technology, the researchers constructed a vector with a gata1 promoter that drives the expression of Cas9 and specifically knocks out the urod gene involved in the synthesis of red blood cell heme. By targeting the Urod gene, the researchers observed red fluorescent red blood cells in zebrafish embryos and repeated the phenotype observed in yquem mutant zebrafish. Mosaic gene disruption occurs in F0 embryos, and this phenotype is very obvious in F1 zebrafish.

　　This CRISPR-based vector system provides a convenient way to achieve tissue-specific gene knockout in zebrafish. This greatly expands the scope of research on zebrafish loss of function and is very important.