CRISPR/Cas9 Gene Disruption Studies in F0 Xenopus Tadpoles: Understanding Development and Disease in the Frog.
CRISPR/Cas9 Gene Disruption Studies in F0 Xenopus Tadpoles: Understanding Development and Disease in the Frog.
复制标题
F0 级爪蟾蝌蚪中的 CRISPR/Cas9 基因破坏研究：了解青蛙的发育和疾病。
DOI：
10.1007/978-1-0716-3004-4_10
复制
发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Abu-Daya A
中科院分区：
文献类型：
--
作者：
Abu-Daya A
CRISPR/Cas9由于其易用性和多功能性，已成为一系列脊椎动物模型生物中基因敲除的最佳方法。基因特异性引导RNA可以被设计为独特的基因组序列并用于靶向Cas9内切核酸酶，其在所需基因座处引起双链断裂。通过非同源末端连接修复断裂通常会导致几个核苷酸的缺失或插入，这通常会导致无义突变。长期以来，Xenopusfrogs一直是研究基因功能的优秀模式生物，它们已被证明在基因编辑实验中很有用，特别是二倍体物种X。热带植物在这一章中，我们提出了我们的协议，基因中断在非洲爪蟾，我们经常使用的研究发展过程和模型人类遗传疾病。
CRISPR/Cas9 has become the favorite method for gene knockouts in a range of vertebrate model organisms due to its ease of use and versatility. Gene-specific guide RNAs can be designed to a unique genomic sequence and used to target the Cas9 endonuclease, which causes a double-stranded break at the desired locus. Repair of the breaks through non-homologous end joining often results in the deletion or insertion of several nucleotides, which frequently result in nonsense mutations.Xenopusfrogs have long been an excellent model organism in which to study gene function, and they have proven to be useful in gene-editing experiments, especially the diploid species,X. tropicalis. In this chapter, we present our protocols for gene disruption inXenopus, which we regularly use to investigate developmental processes and model human genetic disease.