Laboratorio de Desenvolvimento de Vacinas-Centro de Biotecnologia, Instituto Butantan, Sao Paulo, Brazil.
Methods Mol Biol. 2020;2134:109-122. doi: 10.1007/978-1-0716-0459-5_10.
Easy, practical, and affordable gene silencing techniques are constantly progressing, and genetic tools such as TALEs, RNAi, and CRISPR/Cas9 have emerged as new techniques for understanding the basic biology and virulence mechanisms of pathogenic organisms, including bacteria. Here, we describe one-step targeted gene silencing in Leptospira biflexa by using plasmids expressing catalytically inactive Streptococcus pyogenes Cas9 (dCas9) and a single-guide RNA (sgRNA) capable of pairing to the coding strand of a desired gene.
简单、实用且价格合理的基因沉默技术不断发展，TALEs、RNAi 和 CRISPR/Cas9 等遗传工具已成为研究包括细菌在内的病原生物基础生物学和毒力机制的新技术。在这里，我们描述了通过表达无酶活性的酿脓链球菌 Cas9（dCas9）和能够与靶基因编码链配对的单链向导 RNA（sgRNA）的质粒一步靶向性基因沉默在双曲钩端螺旋体中的应用。
