Construction of a cloning vector appropriate for shRNA expression study

Objective:To establish a novel vector with low non-recombinant background and high success rate of cloning and thus suitable for shRNA expression study.Methods:The DNA sequence containing ccdB suicide gene and kanamycin resistance gene was amplified by using PCR and cloned into the shRNA expression plasmid pSilencer3.1-H1 neo that was previously treated with appropriate restriction endonucleases.The power of the resulting vector pSilencer3.1-ccdB was evaluated by cloning the luciferase shRNA expression sequence.As control,the original parent vector pSilencer3.1-H1 was also employed to clone the sequence.Results:The pSilencer3.1-ccdB vector dramatically increased the success rate of cloning by removing the non-recombinant background as compared with the control vector.Conclusion:This study provides a useful tool that will prove its efficacy in the studies of gene function using RNA interference techniques.