Lamarcq Laurence H, Scherer Bradley J, Phelan Michael L, Kalnine Nikolai N, Nguyen Yen H, Kabakova Tatyana, Chen Xiaoyi, Tan Marcia, Chang Cynthia, Berlon Charina, Campos-Gonzalez Roberto, Gao Guo-Jian, Golz Stefan, Vysotski Eugene S, Farmer Andrew A
Clontech Laboratories, Mountain View, CA 94043, USA.
J Biomol Screen. 2006 Apr;11(3):236-46. doi: 10.1177/1087057105284342. Epub 2006 Feb 20.
A method for high-throughput cloning and analysis of short hairpin RNAs (shRNAs) is described. Using this approach, 464 shRNAs against 116 different genes were screened for knockdown efficacy, enabling rapid identification of effective shRNAs against 74 genes. Statistical analysis of the effects of various criteria on the activity of the shRNAs confirmed that some of the rules thought to govern small interfering RNA (siRNA) activity also apply to shRNAs. These include moderate GC content, absence of internal hairpins, and asymmetric thermal stability. However, the authors did not find strong support for position specific rules. In addition, analysis of the data suggests that not all genes are equally susceptible to RNA interference (RNAi).
本文描述了一种用于高通量克隆和分析短发夹RNA（shRNA）的方法。利用这种方法，针对116个不同基因的464个shRNA进行了敲低效率筛选，从而能够快速鉴定出针对74个基因的有效shRNA。对各种标准对shRNA活性影响的统计分析证实，一些被认为支配小干扰RNA（siRNA）活性的规则也适用于shRNA。这些规则包括适度的GC含量、不存在内部发夹结构以及不对称的热稳定性。然而，作者并未找到对位置特异性规则的有力支持。此外，数据分析表明并非所有基因对RNA干扰（RNAi）都同样敏感。
