Short hairpin RNA-expressing bacteria elicit RNA interference in mammals

• Published in: Nature biotechnology Vol. 24; no. 6; pp. 697 - 702
• Main Authors: Li, Chiang J, Xiang, Shuanglin, Fruehauf, Johannes
• Format: Journal Article
• Language: English
• Published: New York, NY Nature 01.06.2006; Nature Publishing Group
• Subjects: Animals; Bacteria; Biological and medical sciences; Biotechnology; Cell Line, Tumor; Colonic Neoplasms - genetics; Colonic Neoplasms - microbiology; E coli; Escherichia coli; Escherichia coli - genetics; Female; Fundamental and applied biological sciences. Psychology; Gene expression; Genomics; Health. Pharmaceutical industry; Humans; Industrial applications and implications. Economical aspects; Intravenous administration; Mammals; Mice; Mice, Inbred C57BL; Mice, Nude; MicroRNAs - genetics; Miscellaneous; Ribonucleic acid; RNA; RNA Interference; RNA, Bacterial - genetics; RNA, Small Interfering - genetics; Gene silencing; Vertebrata; Mammalia; RNA interference; Mouse; Escherichia coli; Rodentia; Bacteria; Gene therapy; Genetically modified microorganism; Enterobacteriaceae
• ISSN: 1087-0156; 1546-1696
• DOI: 10.1038/nbt1211

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 β-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.