Construction and verification of Ientiviral CRKL gene RNA interfering vector / 国际肿瘤学杂志

ABSTRACT

Objective To construct a lentiviral vector carrying CRKL gene RNA interfering( RNAi ).Methods The CRKL RNAi was selected and subcloned into the lentiviral vector,pGCL-GFP(including U6 promotor and green fluorescent protein),generating the lentiviral vector LV-shCRKL.The corrected CRKL was confirmed by endoenzyme digestion ,sequencing.Recombinant lentiviruses were produced by 293T cells following the eo-transfection of LV-shCRKL,with the packaging plasmids pHelper1.0 and pHelper2.0.The virus titer was detected by GFP expressions in 293T cells.Results Plasmid LV-shCRKL carried the correct sequence.The recombinant lentiviruse LV-shCRKL could be produced by co-transfection of LV-shCRKL to 293T cells.Conclusion The recombinant lentiviruse vector LV-shCRKL is constructed successful.