CRISPR/Cas9 plant genome multi-target editing vector and application thereof

• Main Authors: NIU LIFANG, ZHANG PENGCHENG, XIA XIUZHI, LIN HAO, ZHU BUTUO, SUN SHIXU
• Format: Patent
• Language: Chinese; English
• Published: 18.06.2024
• 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 a CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 plant genome multi-target editing vector and application thereof. The number of the sgRNA expression cassettes of the editing vector is two or more than two, and the sgRNA expression cassettes are connected in series; the sgRNA expression cassette is driven to express by a plant endogenous U6 promoter; in combination with a tRNA system, each sgRNA expression cassette can be connected in series with two or more than two target sequence sgRNAs targeting plant genes. The invention further provides a CRISPR/Cas9 multi-target editing vector for editing the medicago sativa MsTFL1s gene, the CRISPR/Cas9 multi-target editing vector comprises three sgRNA expression cassettes which are connected in series, three high-expression medicago sativa endogenous U6 promoters drive and express nine sgRNA target sequences targeting the medicago sativa MsTFL1s gene which are connected in series based on a tRNA-sgRNA system, a vector