Instituto Carlos Chagas, Fiocruz Paraná, Curitiba, Paraná 81350-010, Brazil.
Université de Strasbourg, CNRS, INSERM, Institut de Génétique et de Biologie Moléculaire et Cellulaire, UMR 7104, U 1258, 67404 Illkirch, France.
STAR Protoc. 2022 Apr 21;3(2):101324. doi: 10.1016/j.xpro.2022.101324. eCollection 2022 Jun 17.
This protocol outlines a new genetic complementation strategy to investigate gene function in , the parasite causing Chagas disease. We combine CRISPR-Cas9 technology with recombination of variants of the target gene containing the desired mutations that are resistant to Cas9-cleavage, which enables detailed investigation of protein function. This experimental strategy overcomes some of the limitations associated with gene knockouts in . For complete details on the use and execution of this protocol, please refer to Marek et al. (2021).
本方案概述了一种新的遗传互补策略，用于研究导致恰加斯病的寄生虫 中的基因功能。我们将 CRISPR-Cas9 技术与包含所需突变的靶基因变体的重组相结合，这些突变对 Cas9 切割具有抗性，从而能够详细研究蛋白质功能。这种实验策略克服了 在 中基因敲除相关的一些限制。有关该方案使用和执行的完整详细信息，请参阅 Marek 等人。（2021 年）。
