Department of Pediatrics, the First Affiliated Hospital of Zhengzhou University. Zhengzhou 450052, Henan, People's Republic of China.
J Biochem Mol Toxicol. 2011 Mar-Apr;25(2):55-9. doi: 10.1002/jbt.20331. Epub 2011 Mar 11.
RNA interference (RNAi), an evolutionarily conserved process of gene silencing, is now a common tool in gene functional studies. However, potential "off-target effects" is one of major concerns in RNAi experiment associated with false positive results. Apart from continuing improvement in small interfering RNA (siRNA) designs, there is no method available to prevent the generation of "off-target effects" resulted from possible identity between siRNA and abundant cellular mRNA transcripts. In the present study, we have developed a lentiviral vector-mediated system that allows efficient siRNA silencing and concurrent rescue of targeted genes. While this approach does not eliminate potential "off-target effects," concurrent rescue of a target gene allows a definite judgment with regard to a phenotype change, either from expected siRNA silencing or "off-target effects." The system has been validated with murine CIN85 gene and may be generally applicable in molecular studies from broad fields.
RNA 干扰（RNAi）是一种进化上保守的基因沉默过程，现在是基因功能研究中常用的工具。然而，与假阳性结果相关的 RNAi 实验中，潜在的“脱靶效应”是主要关注点之一。除了不断改进小干扰 RNA（siRNA）设计外，没有方法可以防止 siRNA 与丰富的细胞 mRNA 转录本之间可能存在的同一性导致的“脱靶效应”的产生。在本研究中，我们开发了一种基于慢病毒载体的系统，该系统允许高效的 siRNA 沉默和靶向基因的同时挽救。虽然这种方法不能消除潜在的“脱靶效应”，但目标基因的同时挽救允许对表型变化做出明确判断，无论是来自预期的 siRNA 沉默还是“脱靶效应”。该系统已在鼠 CIN85 基因上得到验证，可能适用于广泛领域的分子研究。
