Kuhlmann Markus, Popova Blagovesta, Nellen Wolfgang
Abt. Genetik, Kassel University, Germany.
Methods Mol Biol. 2006;346:211-26. doi: 10.1385/1-59745-144-4:211.
Knockouts by homologous recombination are frequently used to investigate the function of genes in Dictyostelium and other organisms. Antisense-mediated gene silencing and RNA interference (RNAi) are convenient alternatives to reduce gene expression to different levels and to silence multigene families. We describe here the methods for efficient RNA interference in Dictyostelium and some useful mutant strains that enhance the success rate or may serve as convenient controls. We believe that it is helpful to also discuss failed attempts to optimize and expand the system because these are rarely discussed in the literature. In addition, a list of in silico and experimentally identified components in the RNAi and antisense pathway is presented.
通过同源重组进行基因敲除常用于研究盘基网柄菌和其他生物中基因的功能。反义介导的基因沉默和RNA干扰（RNAi）是将基因表达降低到不同水平以及使多基因家族沉默的便捷替代方法。我们在此描述了在盘基网柄菌中进行高效RNA干扰的方法以及一些有助于提高成功率或可作为便捷对照的有用突变株。我们认为讨论优化和扩展该系统的失败尝试也很有帮助，因为这些在文献中很少被讨论。此外，还列出了RNAi和反义途径中通过计算机模拟和实验鉴定的成分清单。
