Short hairpin RNA-expressing bacteria elicit RNA interference in mammals
Short hairpin RNA-expressing bacteria elicit RNA interference in mammals
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DOI：
10.1038/nbt1211
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发表时间：
2006-06-01
影响因子：
46.9
通讯作者：
Li, Chiang J.
中科院分区：
文献类型：
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作者：
Xiang, Shuanglin;Fruehauf, Johannes;Li, Chiang J.
RNA干扰（RNAi）是一种有效的机制，从植物到人类，用于特异性沉默基因，这为功能基因组学和基因靶向治疗带来了希望。在这里，我们表明，细菌工程产生短发夹RNA（shRNA）靶向哺乳动物基因诱导跨界RNAi在体外和体内。将非致病性大肠杆菌工程化以从含有侵袭素基因Inv和嗜酸乳杆菌溶血素O基因HlyA的质粒转录shRNA，所述侵袭素基因Inv和嗜酸乳杆菌溶血素O基因HlyA编码将shRNA成功转移到哺乳动物细胞中所需的两种细菌因子。口服或静脉给药后，E.大肠杆菌编码针对CTNNB 1（连环蛋白β-1）的shRNA在小鼠肠上皮和人结肠癌异种移植物中诱导显著的基因沉默。这些结果提供了高等生物中跨界RNAi的一个例子，并表明细菌介导的RNAi在功能基因组学、治疗靶点验证和开发临床相容的基于RNAi的疗法方面的潜力。
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.