Anyone tried to express a transgene from a native 5’LTR promoter of the lentiviral system. Literature says that 5' LTR promoter is inactivated upon integration into the target genome to prevent the replication of the viral sequences.
My transgenes are quite large (total viral genome from 5’ LTR to 3’ LTR: 5-11 kb with introns, and 5-9 kb upon splicing). I am not able to produce any functional viral particles above 5 kb size. Hence exploring on the possibility to reduce the overall genome size. In this regard, I am thinking of removing the CMV promoter used to drive the transgene and then relay on the 5’ LTR promoter to drive the transgene.
Any suggestions in this regard would be highly appreciated.
Many thanks
Manjunatha