Rapid selection of accessible and cleavable sites in RNA by Escherichia coli RNase P and random external guide sequences
- *Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, CT 06520; and
- †Reference Center for Detection of Antimicrobial Resistance (K-res), Department of Microbiology and Infection Control, University Hospital of North Norway, 9038 Tromso, Norway
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Contributed by Sidney Altman, December 20, 2007 (received for review November 16, 2007)
Abstract
A method of inhibiting the expression of particular genes by using external guide sequences (EGSs) has been improved in its rapidity and specificity. Random EGSs that have 14-nt random sequences are used in the selection procedure for an EGS that attacks the mRNA for a gene in a particular location. A mixture of the random EGSs, the particular target RNA, and RNase P is used in the diagnostic procedure, which, after completion, is analyzed in a gel with suitable control lanes. Within a few hours, the procedure is complete. The action of EGSs designed by an older method is compared with EGSs designed by the random EGS method on mRNAs from two bacterial pathogens.
Footnotes
- ‡To whom correspondence should be addressed. E-mail: sidney.altman{at}yale.edu
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Author contributions: G.X. and J.-h.K. contributed equally to this work; E.W.L., G.X., J.-h.K., designed research; E.W.L., G.X., and J.-h.K performed research; E.W.L., G.X., J.-h.K., and S.A. analyzed data; and E.W.L., G.X., J.-h.K., and S.A. wrote the paper.
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The authors declare no conflict of interest.
- © 2008 by The National Academy of Sciences of the USA
