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Persistence of seed-based activity following microRNA segmentation


ABSTRACT: microRNAs are M-bM-^HM-<22 nucleotide regulatory RNAs that are processed into duplexes from hairpin structures and incorporated into Argonaute proteins. We find that a nick in the middle of the guide strand of an miRNA sequence allows for seed-based targeting characteristic of miRNA activity. Insertion of an inverted abasic, a dye, or a small gap between the two segments still permits target knockdown. While activity from the seed region of the segmented miRNA is apparent, activity from the 3' half of the guide strand is impaired, suggesting that an intact guide backbone is required for contribution from the 3' half. miRNA activity was also observed following nicking of a miRNA precursor. These results illustrate a structural flexibility in miRNA duplexes and may have applications in the design of miRNA mimetics. For the microarray portion of the paper, a miR-124 duplex containing a segmented guide strand (G10.12/P) or an intact guide strand (G/P) was transfected into HCT-116 dicer- cells. Profiles were compared with mock transfections.

ORGANISM(S): Homo sapiens

SUBMITTER: Lee Lim 

PROVIDER: E-GEOD-25224 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Persistence of seed-based activity following segmentation of a microRNA guide strand.

Chorn Guillaume G   Zhao Lihong L   Sachs Alan B AB   Flanagan W Michael WM   Lim Lee P LP  

RNA (New York, N.Y.) 20101022 12


microRNAs are ∼ 22 nucleotide regulatory RNAs that are processed into duplexes from hairpin structures and incorporated into Argonaute proteins. Here, we show that a nick in the middle of the guide strand of an miRNA sequence allows for seed-based targeting characteristic of miRNA activity. Insertion of an inverted abasic, a dye, or a small gap between the two segments still permits target knockdown. While activity from the seed region of the segmented miRNA is apparent, activity from the 3' hal  ...[more]

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