Method for increasing targeting force of NK cells and application of CCR5 gene
A technology of NK cells, 1. CCR5, applied in the biological field, can solve the problems of restricting NK cell therapy and weak directional movement of NK cells, and achieve strong tumor infiltration ability, good clinical transformation prospects, and the effect of inhibiting tumor growth.
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[0023] The experimental methods in the following examples are conventional methods unless otherwise specified, and the involved experimental reagents and materials are conventional biochemical reagents and materials unless otherwise specified.
[0024] 1. Acquisition of CCR5 coding sequence
[0025] After the total RNA of human peripheral blood mononuclear cells was extracted with Trizol reagent (Invitrogen), it was reverse transcribed into cDNA. Then, PCR amplification obtains the coding sequence of CCR5 (as shown in SEQ ID NO.1), wherein upstream primer sequence: 5'- ggatcc atggattatcaagtgtcaag-3' (the underline is the BamHI restriction site), downstream primer sequence: 5'- gcggccgc caagccccacagatatttc-3' (the underline is the NotI restriction site).
[0026] 2. Construction of CCR5 lentiviral overexpression plasmid
[0027] The PCR product of lentiviral plasmid pCDH-EF1-MSC-T2A-conGFP or CCR5 coding sequence was double digested with BamHI and NotI, respectively. After...
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