Off-Target Effect of doublecortin Family shRNA on Neuronal Migration Associated with Endogenous MicroRNA Dysregulation
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SCOPUS
- Title
- Off-Target Effect of doublecortin Family shRNA on Neuronal Migration Associated with Endogenous MicroRNA Dysregulation
- Authors
- Baek, ST; Kerjan, G; Bielas, SL; Lee, JE; Fenstermaker, AG; Novarino, G; Gleeson, JG
- Date Issued
- 2014-06-18
- Publisher
- CELL PRESS
- Abstract
- Acute gene inactivation using short hairpin RNA (shRNA, knockdown) in developing brain is a powerful technique to study genetic function; however, discrepancies between knockdown and knockout murine phenotypes have left unanswered questions. For example, doublecortin (Dcx) knockdown but not knockout shows a neocortical neuronal migration phenotype. Here we report that in utero electroporation of shRNA, but not siRNA or miRNA, to Dcx demonstrates a migration phenotype in Dcx knockouts akin to the effect in wild-type mice, suggesting shRNA-mediated off-target toxicity. This effect was not limited to Dcx, as it was observed in Dclk1 knockouts, as well as with a fraction of scrambled shRNAs, suggesting a sequence-dependent but not sequence-specific effect. Profiling RNAs from electroporated cells showed a defect in endogenous let7 miRNA levels, and disruption of let7 or Dicer recapitulated the migration defect. The results suggest that shRNA-mediated knockdown can produce untoward migration effects by altering endogenous miRNA pathways.
- URI
- https://oasis.postech.ac.kr/handle/2014.oak/37088
- DOI
- 10.1016/J.NEURON.2014.04.036
- ISSN
- 0896-6273
- Article Type
- Article
- Citation
- NEURON, vol. 82, no. 6, page. 1255 - 1262, 2014-06-18
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- There are no files associated with this item.
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