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| Status |
Public on Nov 12, 2010 |
| Title |
Persistence of seed-based activity following microRNA segmentation |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
|
| Summary |
microRNAs are ∼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.
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| Overall design |
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.
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| |
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| Contributor(s) |
Chorn G, Zhao L, Sachs A, Flanagan M, Lim L |
| Citation(s) |
20971811 |
| Submission date |
Nov 09, 2010 |
| Last update date |
Jul 09, 2014 |
| Contact name |
Lee Lim |
| E-mail(s) |
lee_lim@merck.com
|
| Phone |
206-802-7323
|
| Organization name |
Rosetta Inpharmatics
|
| Street address |
|
| City |
Seattle |
| State/province |
WA |
| ZIP/Postal code |
98109 |
| Country |
USA |
| |
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| Platforms (1) |
| GPL10379 |
Rosetta/Merck Human RSTA Custom Affymetrix 2.0 microarray [HuRSTA-2a520709] |
|
| Samples (4)
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| GSM619260 |
Control #1: mock transfection of HCT116 dicer- cell line, 24 h timepoint |
| GSM620811 |
Control #2: mock transfection of HCT116 dicer- cell line, 24 h timepoint |
| GSM620812 |
Transfection of HCT116 dicer- cell line with miR-124 G/P, 24h timepoint |
| GSM620813 |
Transfection of HCT116 dicer- cell line with miR-124 G10.12/P, 24h timepoint |
|
| Relations |
| BioProject |
PRJNA134463 |
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