589. Short Hairpin RNAs Based on miR-451 Show Potent Knockdown without Passenger Strand Activity

TY - ABST

T1 - 589. Short Hairpin RNAs Based on miR-451 Show Potent Knockdown without Passenger Strand Activity

AU - Kaadt, Erik

AU - Aagaard, Lars

PY - 2016/5/1

Y1 - 2016/5/1

N2 - Based on the recent findings that RNAi designs with shortened shRNA stems bypass Dicer cleavage and rely on Ago2 for processing of the ‘effector’ guide strand, we have compared designs based on ‘classical’ shRNA structures with shortened stems and mimics of miR-451. Our results support the complete lack of passenger stands activity which is likely to improve safety in therapeutic applications. We report that miR-451 mimics are more potent than shortened shRNAs and our data further suggests that the U6 promoter is the preferred choice as compared to H1. We suggest that imprecise transcription initiation from H1 promoter makes it hard to predict the actual target site(s) and may thus be a safety concern. Tiling of U6-promoted miR-451 mimics supports this notion and demonstrate that knockdown efficacy vary substantially when shifting the target site one nucleotide.

AB - Based on the recent findings that RNAi designs with shortened shRNA stems bypass Dicer cleavage and rely on Ago2 for processing of the ‘effector’ guide strand, we have compared designs based on ‘classical’ shRNA structures with shortened stems and mimics of miR-451. Our results support the complete lack of passenger stands activity which is likely to improve safety in therapeutic applications. We report that miR-451 mimics are more potent than shortened shRNAs and our data further suggests that the U6 promoter is the preferred choice as compared to H1. We suggest that imprecise transcription initiation from H1 promoter makes it hard to predict the actual target site(s) and may thus be a safety concern. Tiling of U6-promoted miR-451 mimics supports this notion and demonstrate that knockdown efficacy vary substantially when shifting the target site one nucleotide.

UR - https://www.cell.com/molecular-therapy-family/molecular-therapy/comments/S1525-0016(16)33397-4

M3 - Conference abstract for conference

SP - S233-S234

ER -