Construction of the Recombinant Lentivirus of RNAi Targeting Human EGFL7 Gene.

WANG Ai-yue; LI Qiang; XU Qiong-guan; LIU Da-yuan; XU Peng-xiang; YU Dan

Objective: To construct the recombinant lentivirus of RNAi targeting human Epidermal Growth Factor-like domain 7 (EGFL7) gene. Methods: The RNAi sequences targeting human EGFL7 gene were inserted into pLV3 lentivirus vectors containing green fluorescent protein. Recombinant plasmids were thus obtained and transfected to 293T cells along with packaging plasmids pRsv-REV, pMDlg-pRRE and pMD2G, so as to produce recombinant lentivirus, which later infected HUVECs. The gene silencing effect was then determined through examination of the resulting mRNA level of the targeted gene in the HUVECs. Results: Three RNAi sequences of human EGFL7 gene were selected out and packaged into recombinant lentivirus, with the titers of 1× 108, 2× 108 and 5× 108 TU/mL, respectively. The interfering effect of each sequence was assessed and compared. Conclusions: Three effective RNAi targeting sequences for human EGFL7 gene were selected out, and recombinant lentivirus was successfully constructed. It was demonstrated that the sequences can significantly silence the expression of the EGFL7 gene in HUVECs.

RECOMBINANT viruses; LENTIVIRUSES; RNA interference

Progress in Modern Biomedicine, 2016, Vol 16, Issue 26, p5042

1673-6273

Academic Journal

10.13241/j.cnki.pmb.2016.26.010