question: RNAi for bacteria? - (Feb/06/2012 )

Hi, I want to silence a gene in a G(+) bacteria. I want to ask will siRNA or shRNA work for prokaryotes? If not, what technique can I use?

Thanks a lot!

Here's one approach - Morpholino antisense conjugated with cell-penetrating peptides. The peptide conjugates aren't commercially available, but the conjugation is fairly straightforward and protocols have been published.

Greenberg DE, Marshall-Batty KR, Brinster LR, Zarember KA, Shaw PA, Mellbye BL, Iversen PL, Holland SM, Geller BL. Antisense Phosphorodiamidate Morpholino Oligomers Targeted to an Essential Gene Inhibit Burkholderia cepacia Complex. J Infect Dis. 2010 May 3.

Mellbye BL, Weller DD, Hassinger JN, Reeves MD, Lovejoy CE, Iversen PL, Geller BL. Cationic phosphorodiamidate morpholino oligomers efficiently prevent growth of Escherichia coli in vitro and in vivo. J Antimicrob Chemother. 2009 Nov 1.

Mitev GM, Mellbye BL, Iversen PL, Geller BL. Inhibition of Intracellular Growth of Salmonella enterica serovar typhimurium in Tissue Culture by Antisense Peptide-Phosphorodiamidate Morpholino Oligomer. Antimicrob Agents Chemother. 2009 Jul 6.

Shen N, Ko JH, Xiao G, Wesolowski D, Shan G, Geller B, Izadjoo M, Altman S. Inactivation of expression of several genes in a variety of bacterial species by EGS technology. Proc Natl Acad Sci U S A. 2009 Apr 30.

Mellbye BL, Puckett SE, Tilley LD, Iversen PL, Geller BL. Variations in Amino Acid Composition of Antisense Peptide-Phosphorodiamidate Morpholino Oligomer Affect Potency Against Escherichia coli in vitro and in vivo. Antimicrob Agents Chemother. 2009 Feb;53(2):525-30. Epub 2008 Nov 17.

Tilley LD, Mellbye BL, Puckett SE, Iversen PL, Geller BL. Antisense peptide-phosphorodiamidate morpholino oligomer conjugate: dose-response in mice infected with Escherichia coli. J Antimicrob Chemother. 2007 Jan;59(1):66-73. Epub 2006 Oct 31.

Tilley LD, Hine OS, Kellogg JA, Hassinger JN, Weller DD, Iversen PL, Geller BL. Gene-Specific Effects of Antisense Phosphorodiamidate Morpholino Oligomer-Peptide Conjugates on Escherichia coli and Salmonella enterica Serovar Typhimurium in Pure Culture and in Tissue Culture. Antimicrob Agents Chemother. 2006 Aug;50(8):2789-96.

Deere J, Iversen P, Geller BL. Antisense Phosphorodiamidate Morpholino Oligomer Length and Target Position Effects on Gene-Specific Inhibition in Escherichia coli. Antimicrob Agents Chemother. 2005 Jan;49(1):249-55.

Geller BL, Deere J, Tilley L, Iversen PL. Antisense phosphorodiamidate morpholino oligomer inhibits viability of Escherichia coli in pure culture and in mouse peritonitis. J Antimicrob Chemother. 2005 Jun;55(6):983-8. Epub 2005 May 4.

Geller BL. Antibacterial antisense. Curr Opin Mol Ther 2005 7(2):109-13.

Geller BL, Deere JD, Stein DA, Kroeker AD, Moulton HM, Iversen PL. Inhibition of Gene Expression in Escherichia coli by Antisense Phosphorodiamidate Morpholino Oligomers. Antimicrob Agents Chemother. 2003 Oct;47(10):3233-9.

Hi! Thanks very much for your suggestions!

A bit more experimental: There is in some bacteria a gene active called hfq, if they are active in hfq you can try to use it for some knockdown similar to shRNAs.
http://mcb.illinois.edu/faculty/profile/cvanderp