Title |
Improved siRNA/shRNA Functionality by Mismatched Duplex
|
---|---|
Published in |
PLOS ONE, December 2011
|
DOI | 10.1371/journal.pone.0028580 |
Pubmed ID | |
Authors |
Haoquan Wu, Hongming Ma, Chunting Ye, Danielle Ramirez, Shuiping Chen, Jessica Montoya, Premlata Shankar, Xiaozhong A. Wang, N. Manjunath |
Abstract |
siRNA (small interfering RNA) and shRNA (small hairpin RNA) are powerful and commonly used tools in biomedical research. Currently, siRNAs are generally designed as two 21 nt strands of RNA that include a 19 nt completely complementary part and a 2 nt overhang. However, since the si/shRNAs use the endogenous miRNA machinery for gene silencing and the miRNAs are generally 22 nt in length and contain multiple internal mismatches, we tested if the functionality can be increased by designing the si/shRNAs to mimic a miRNA structure. We systematically investigated the effect of single or multiple mismatches introduced in the passenger strand at different positions on siRNA functionality. Mismatches at certain positions could significantly increase the functionality of siRNAs and also, in some cases decreased the unwanted passenger strand functionality. The same strategy could also be used to design shRNAs. Finally, we showed that both si and miRNA structured oligos (siRNA with or without mismatches in the passenger strand) can repress targets in all individual Ago containing cells, suggesting that the Ago proteins do not differentiate between si/miRNA-based structure for silencing activity. |
X Demographics
Geographical breakdown
Country | Count | As % |
---|---|---|
United States | 1 | 50% |
Australia | 1 | 50% |
Demographic breakdown
Type | Count | As % |
---|---|---|
Members of the public | 1 | 50% |
Scientists | 1 | 50% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Japan | 1 | 1% |
France | 1 | 1% |
Germany | 1 | 1% |
Austria | 1 | 1% |
Unknown | 65 | 94% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 15 | 22% |
Researcher | 15 | 22% |
Student > Master | 10 | 14% |
Student > Doctoral Student | 4 | 6% |
Other | 4 | 6% |
Other | 12 | 17% |
Unknown | 9 | 13% |
Readers by discipline | Count | As % |
---|---|---|
Agricultural and Biological Sciences | 32 | 46% |
Biochemistry, Genetics and Molecular Biology | 15 | 22% |
Medicine and Dentistry | 5 | 7% |
Chemistry | 3 | 4% |
Immunology and Microbiology | 2 | 3% |
Other | 3 | 4% |
Unknown | 9 | 13% |