Secondary structure of target RNA impacts on RNA interference efficiency / 基础医学与临床

ABSTRACT

Objective To study the effects of the RNA secondary structure on the RNA interference efficacy and to optimize the RNAi target sequence selection.Methods N-Ras mRNA in K562 human chronic erythro-leukemia cell line was selected as the target for RNAi experiments.Four siRNAs were designed aiming at secondary structure region and none-secondary structure region separately.The N-Ras expression change and the cell growth was tested by semi-quantitative reverse transcription polymerase chain reaction(RT-PCR),light microscopy,MTT,and flow cytometry.Results Knock-down efficiency was recorded in all of the 4 experiment groups,but the degree varied a lot.The siRNAs targeting none-secondary structure region presented higher silencing efficiency than those targeting secondary structure region.Conclusion The secondary structure of mRNA closely relate to the RNA interference efficiency,which should be considered in the processing of the target sequence selection.