I am trying to modify a retroviral vector PMIG II which has 5' LTR and 3' LTR used as promoters. The 5'LTR is followed by multiple cloning site (MCS). I am looking to clone my gene of interest under granzyme B mouse promoter at MCS site. Do I need to remove 5'LTR and 3'LTR in the vector in order to do that? Can 5'LTR drive my granzyme B Promoter?. The vector would be ultimately used to transduce mouse cell line.
Thanks in advance for the feedback.
Dear Preeti Thakur
It is common to insert another promoter between the 5'LTR and 3'LTR of the Lenti vector to control downstream gene expression. The use of the granzyme B promoter is expected to result in activated T cell-specific gene expression, but in order to make it more specific, it is recommended to use SIN vectors with reduced activity of the 5'LTR and 3'LTR. Even so, it may be difficult to achieve completely specific expression, so it may be necessary to devise some method as described in the following paper.
https://cancerres.aacrjournals.org/content/canres/70/24/10141.full.pdf
@Narumi Uno Thank you so much for your feedback. I really appreciate it!. I would look at this strategy and design accordingly.