Construction method and application of WNT1 mutant gene knock-in rat osteogenesis imperfect animal model

• Main Authors: XU HONGWEN, WANG XIUMIN, TANG YAPING, SHANG LIYUAN, ZHU MINGWEI, LI XIN, SHI WEIZHE, XU YIBO, NONG TIANYING, LI XIA, XIAN CAIXIA
• Format: Patent
• Language: Chinese; English
• Published: 21.06.2024
• Subjects: AGRICULTURE; ANIMAL HUSBANDRY; BEER; BIOCHEMISTRY; CARE OF BIRDS, FISHES, INSECTS; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; FISHING; FORESTRY; HUMAN NECESSITIES; HUNTING; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; NEW BREEDS OF ANIMALS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; SPIRITS; TRAPPING; VINEGAR; WINE

The invention belongs to the technical field of biomedicine, discloses a construction method and application of a WNT1 mutant gene knock-in rat osteogenesis imperfect animal model, and particularly discloses sgRNA of a specific target WNT1 mutant gene, and the sequence of the sgRNA is GCCGATGGTGGTAAGTGAGCTGG. Or, ACCAGCTCACTTACCACCATGG, or, the mutation site of the WNT1 mutant gene is c.104 + 1Ggt; a. According to the sgRNA provided by the invention, a WNT1 mutant gene (c.104 + 1Ggt) can be constructed on the basis of a CRISPR/Cas9 gene editing technology; the method comprises the following steps: A) knocking into an animal model for screening medicines for osteogenesis imperfecta or researching the pathogenic mechanism of osteogenesis imperfecta; 本发明属于生物医学技术领域，公开了一种WNT1突变基因敲入大鼠成骨不全动物模型的构建方法和应用，具体公开了一种特异性靶向WNT1突变基因的sgRNA，所述sgRNA的序列为GCCGATGGTGGTAAGTGAGCTGG；或ACCAGCTCACTTACCACCATCGG；所述WNT1突变基因的突变位点为c.104+1G>A。通过本发明提供的sgRNA，可基于CRISPR/Cas9基因编辑技术构建得到WNT1突变基因(c.104+1G>A)敲入动物模型，用于对成骨不全疾病的药物的筛选或成骨不全疾病的致病机制的研究。