Oligonucleotides Block Small RNA Function

A new study describes a rapid and reliable method for knocking out both types of small regulatory RNA molecules, siRNAs and microRNAs, which allows experimentation to determine how they function.

Investigators at the University of Massachusetts Medical School (Worcester, USA; www.umassmed.edu) worked with a series of models including Drosophila embryo lysates, HeLa cell S100 extracts, cultured human HeLa cells, and Caenorhabditis elegans worms to show that small nucleotide sequences, or 2Œ-O-methyl oligonucleotides, could block the activity of small RNAs.

The highlight of this work, which was published in the February 24, 2004, online edition of PloS, the Public Library of Science, was the finding that C elegans larvae injected with the 2Œ-O-methyl oligonucleotide complementary to the miRNA let-7 developed similarly to larvae lacking the let-7 gene. Let-7 blocks the production of the protein Lin-41 and is important for proper developmental timing in roundworm larvae.

The authors concluded, gWe demonstrate that 2Œ-O-methyl RNA oligonucleotides can provide an efficient and straightforward way to block small RNA function in vivo and furthermore can be used to identify small RNA-associated proteins that mediate RNA silencing pathways.h




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