Pseudotyped HIV was created by removing envelope and adding VSV-G (glycoprotein of vesicular stomatitis virus) also encoding gene for luciferase in pNL4.3. After transfection of cell A and B with plasmid and producing virus, 100 ng/mL virus was used to infect 1 million CD4/mL. After 2 days, luciferase activity in CD4 T cells was shown only by virus produced from cell A, but not from cell B. What type of defects in cell B may be responsible for the defect?
Homa Nath Sharma Is it possible that cell B had poor plasmid uptake during transfection and or the genomic architecture of cell B inhibited the transcription of the HIV genome? What transfection method did you use? You can try and optimize the transfection process by using chemicals such as sodium butyrate that help improve transfection efficiency primarily by enhancing gene expression after DNA uptake.
Sidney T. Sithole Yes that may be possible as I used Myrius TransIT_293 and TransIT-LT1 transfection reagent for plasmid transfection to those cells. Thank you.
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