公开(公告)号：US20250160308A1

公开(公告)日：2025-05-22

申请号：US18840525

申请日：2023-05-17

Applicant: SHANGHAITECH UNIVERSITY

Inventor： Tian CHI ,  Bo LIU ,  Zhengyu JING ,  Xiaoming ZHANG ,  Shaoshuai MAO ,  Yuxin CHEN

IPC: A01K67/0278 ,  C12N9/12 ,  C12N9/22 ,  C12N15/11 ,  C12N15/90

Abstract: Provided are a nucleic acid construct based on a Cre-LoxP recombination system and a CRISPR gene editing system and use thereof. The Cre-LoxP recombination system comprises a Cre enzyme and a LoxP nucleic acid combination. The LoxP nucleic acid combination comprises TATA-Lox71 and TATA-LoxTC9 sequences, which can only be recombined once under the catalysis of the Cre enzyme. The nucleic acid construct carries an inert “stuffer sequence” with a certain length. The nucleic acid construct can express a plurality of sgRNAs in a low bias manner in vivo, but a same cell can only express one sgRNA, thereby efficiently generating mosaicism whose utilities include accurate and sensitive in-situ CRISPR gene screening and rapid, cost-effective preparation of a single-gene knockout line.