Xiang Shuanglin, Fruehauf Johannes, Li Chiang J
Division of Gastroenterology, Beth Israel Deaconess Medical Center, Harvard Medical School, 330 Brookline Ave., Boston, Massachusetts 02215, USA.
Nat Biotechnol. 2006 Jun;24(6):697-702. doi: 10.1038/nbt1211. Epub 2006 May 14.
RNA-interference (RNAi) is a potent mechanism, conserved from plants to humans for specific silencing of genes, which holds promise for functional genomics and gene-targeted therapies. Here we show that bacteria engineered to produce a short hairpin RNA (shRNA) targeting a mammalian gene induce trans-kingdom RNAi in vitro and in vivo. Nonpathogenic Escherichia coli were engineered to transcribe shRNAs from a plasmid containing the invasin gene Inv and the listeriolysin O gene HlyA, which encode two bacterial factors needed for successful transfer of the shRNAs into mammalian cells. Upon oral or intravenous administration, E. coli encoding shRNA against CTNNB1 (catenin beta-1) induce significant gene silencing in the intestinal epithelium and in human colon cancer xenografts in mice. These results provide an example of trans-kingdom RNAi in higher organisms and suggest the potential of bacteria-mediated RNAi for functional genomics, therapeutic target validation and development of clinically compatible RNAi-based therapies.
RNA干扰（RNAi）是一种强大的机制，从植物到人类都保守存在，用于特定基因的沉默，这为功能基因组学和基因靶向治疗带来了希望。在此我们表明，经工程改造能产生靶向哺乳动物基因的短发夹RNA（shRNA）的细菌，可在体外和体内诱导跨物种RNAi。非致病性大肠杆菌经工程改造，可从含有侵袭素基因Inv和溶菌酶O基因HlyA的质粒转录shRNA，这两个基因编码将shRNA成功转移到哺乳动物细胞所需的两种细菌因子。经口服或静脉给药后，编码针对CTNNB1（连环蛋白β-1）的shRNA的大肠杆菌可在小鼠的肠上皮和人结肠癌异种移植瘤中诱导显著的基因沉默。这些结果提供了高等生物中跨物种RNAi的一个实例，并表明细菌介导的RNAi在功能基因组学、治疗靶点验证以及开发临床适用的基于RNAi的疗法方面具有潜力。
