Hannus, Michael and Beitzinger, Michaela and Engelmann, Julia C. and Weickert, Marie-Theresa and Spang, Rainer and Hannus, Stefan and Meister, Gunter (2014) siPools: highly complex but accurately defined siRNA pools eliminate off-target effects. NUCLEIC ACIDS RESEARCH, 42 (12). pp. 8049-8061. ISSN 0305-1048, 1362-4962
Full text not available from this repository. (Request a copy)Abstract
Short interfering RNAs (siRNAs) are widely used as tool for gene inactivation in basic research and therapeutic applications. One of the major shortcomings of siRNA experiments are sequence-specific off-target effects. Such effects are largely unpredictable because siRNAs can affect partially complementary sequences and function like microRNAs (miRNAs), which inhibit gene expression on mRNA stability or translational levels. Here we demonstrate that novel, enzymatically generated siRNA pools-referred to as siPools-containing up to 60 accurately defined siRNAs eliminate off-target effects. This is achieved by the low concentration of each individual siRNA diluting sequence-specific off-target effects below detection limits. In fact, whole transcriptome analyses reveal that single siRNA transfections can severely affect global gene expression. However, when complex siRNA pools are transfected, almost no transcriptome alterations are observed. Taken together, we present enzymatically produced complex but accurately defined siRNA pools with potent on-target silencing but without detectable off-target effects.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | SHORT INTERFERING RNAS; LONG NONCODING RNAS; ARGONAUTE PROTEINS; HUMAN-CELLS; CHEMICAL-MODIFICATION; PASSENGER-STRAND; GUIDE STRAND; CLEAVAGE; SCREENS; BIOINFORMATICS; |
| Subjects: | 500 Science > 570 Life sciences 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Institut für Funktionelle Genomik > Lehrstuhl für Statistische Bioinformatik (Prof. Spang) Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie I > Prof. Dr. Gunter Meister |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 29 Nov 2019 08:17 |
| Last Modified: | 29 Nov 2019 08:17 |
| URI: | https://pred.uni-regensburg.de/id/eprint/10960 |
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