Successful construction of CRISPR/Cas9 Efficient Genome Editing Technology (CAGO) without new gRNA plasmids

CRISPR/Cas9 as a revolutionary genome editing technology has been widely used in a variety of biological genome seditation, this technology in specific applications, such as the presence of "PAM-free" and "CRISPR-tolerant" regions of the genome can not be edited using traditional CRISPR/Cas9 technology; To solve these problems
, the research team of microbial synthetic biotechnology led by Bi Changxuan, a researcher at the Tianjin Institute of Industrial Biotechnology of the Chinese Academy of Sciences, and the research team of microbial metabolism engineering led by researcher Zhang Xueli, have successfully constructed crispR/Cas9 high-efficiency genome editing technology (CAGO) without the need for new gRNA plasmids. First,
, an edited fragment containing a special "N20PAM" sequence is integrated into the genome by homologous recombination, and crispR/Cas9 and red homologous recombination technology is used to achieve homologous recombination within the genome at the insertion site, thus enabling genome indentation.
the method can achieve non-target editing at any site of the genome, the editing process only needs to synthesize an edit fragment, with the help of a common pCAGO plasmid can be edited without the need to build gRNA plasmids.
the construction of this method provides a simple and easy new technical means for synthetic biology research, and lowers the threshold of crispR/Cas9 technology.
research was supported by the National High Technology Research and Development Program (863 Program) and tianjin Key Projects, and the results were published in Scientific Reports.
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