An optimized microRNA scaffold increases shRNA processing and target knockdown

ABSTRACT: Two shRNAs were placed into expression vectors harboring mir30 microRNA scaffold and an optimized scaffold where the artificial restriction sights in mir30 have been removed. After infection and selection shRNA processing was assessed by small-RNA cloning. For both shRNAs, placement into the optimized scaffold resulted in a ~two-fold increase in processing (based on smallRNA levels). Purpose: Others have reported that the EcoRI site that was introduced to the mir30 scaffold results in decreased smallRNA processing and hence reduced target knockdown. We've developed an alternative scaffold (termed ultramir) where this site is removed. smallRNA cloning was used to determine if the movement of this sight resulted in an increase in shRNA processing. Method: Two shRNAs (one targeting Renilla Lu

ORGANISM(S): Gallus gallus

SUBMITTER: Simon Knott 

PROVIDER: E-GEOD-62187 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress