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question: RNAi for bacteria?


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#1 joy123



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Posted 06 February 2012 - 12:20 PM

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!

#2 Jon Moulton

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Posted 07 February 2012 - 08:55 AM

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. [Epub ahead of print]

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. [Epub ahead of print]

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. [Epub ahead of print]

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. [Epub ahead of print]

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.
Jon D. Moulton
Gene Tools, LLC

#3 joy123



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Posted 08 February 2012 - 02:53 PM

Hi! Thanks very much for your suggestions!

#4 wincel



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Posted 17 February 2012 - 01:57 PM

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.

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