Construction of recombinant vectors expressing antisense RNA to CXCR4 and its expression in eukaryotic cells

Objective:To construct recombinant vector expressing antisense RNA to CXCR4 in eukaryotic cells, which will be used to block HIV 1 infection. Methods:The DNA fragment targeted to the initional part of CXCR4 mRNA translation obtained by RT PCR from peripheral blood mononuclear cells (PBMCs) was inserted into retroviral vector pLXSN. The resultant vectors were identified by PCR, restriction endonucleases analyzing and DNA sequencing, then transfected into packaging cells (PA317) with lipofect AMINE. After 2-3 weeks selecting with G418, the pseudovirion in the survival cells′ supernatant was detected by Fluorogenic Quantitative RT PCR, then used to infect NIH/3T3 cells. Results:The expression vector of sense/antisense RNA to CXCR4 had been constructed. The pseudovirion had successfully infected NIH/3T3 cells. The vector had been incorporated into its genome and transcripted into RNA. Conclusion:The gene fragment of antisense RNA to CXCR4 could be obtained from PBMCs and could be transfected into eukaryotic cells with retroviral vector. The results have laid a foundation for studying its inhibiting effect on HIV 1 infection.