SgRNA for editing non-heading Chinese cabbage CENH3 gene and CRISPR/Cas9 vector and application thereof

• Main Authors: HOU XILIN, LIU TONGKUN, HU CHUNMEI, XIAO DONG, LI YING, ZHANG CHANGWEI, WANG JIANJUN, PAN YAOHUA, REN YIMING
• Format: Patent
• Language: Chinese; English
• Published: 26.10.2021
• Subjects: AGRICULTURE; ANIMAL HUSBANDRY; BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; FISHING; FORESTRY; HUMAN NECESSITIES; HUNTING; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; NEW PLANTS OR PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; TRAPPING; VINEGAR; WINE

The invention discloses sgRNA for editing a non-heading Chinese cabbage CENH3 gene and a CRISPR/Cas9 vector and application thereof. According to the invention, the sgRNA of the non-heading Chinese cabbage CENH3 gene is designed, the CRISPR/Cas9 gene editing vector is constructed, the CRISPR/Cas9 vector connected with a target DNA fragment is transferred into a non-heading Chinese cabbage genome, and site-directed mutation of the non-heading Chinese cabbage CENH3 gene is achieved through gene editing. According to the invention, the endogenous gene CENH3 of the non-heading Chinese cabbage is edited by combining a pollen nano magnetic transformation technology with a CRISPR/Cas9 technology, a CENH3 haploid induction system is obtained through plant resistance screening, PCR detection and gene sequencing screening, and a foundation is laid for obtaining haploids by utilizing a CENH3-mediated chromosome elimination mechanism in non-heading Chinese cabbage double haploid breeding. 本发明公开编辑不结球白菜CENH3基因的sgRNA及其CRISP