The invention discloses a CAR-T
transgene vector based on replication defective recombinant
lentivirus. The CAR-T
transgene vector comprises an original nuclear
replicon pUCOri sequence, a resistance
gene AmpR sequence containing
ampicillin, a
virus replicon SV40 Ori sequence, a
lentivirus packaging cis element, ZsGreen1
green fluorescent protein, an IRES
ribosome binding sequence, a human EF1 alpha
promoter , a
chimeric antigen receptor of second-generation CAR or third-generation CAR and a regulating element, wherein the original nuclear
replicon pUCOri sequence is used for
plasmid replication; the resistance
gene AmpR sequence is used for massively proliferating target strains; the
virus replicon SV40 Ori sequence is used for enhancing replication in eukaryocyte; the
lentivirus packaging cis element is used for lentivirus packaging; the ZsGreen1
green fluorescent protein is used for expressing green fluorescent for eukaryocyte; the IRES
ribosome binding sequence is used for jointly transcribing and expressing
protein; the human EF1 alpha
promoter is used for conducting eukaryotic transcription on
antigen receptor genes; the
chimeric antigen receptor is used for forming the second-generation CAR or the third-generation CAR integrating recognition, transfer and start; the regulating element is used for enhancing expression efficiency of transgenes and used after eWPRE-enhanced type woodchuck
hepatitis b
virus is transcribed. Besides, the invention further discloses a construction method and application of the vector. By means of the CAR-T
transgene vector and the construction method and application of the vector,
secretion of
cell factors and an
in vitro killing effect of CAR-T cells can be remarkably improved, and the
clinical treatment effect is remarkable.