Hi! I used lenti-virus system to produce a stable expression cell line. However, cells are able to survive puromycin treatment (lethal concentration for wild type cells) but cannot detect mRNA or protein expression of target gene or protein. The donor plasmid is fine because I checked the expression of the target protein by Lipo2000 transfection. How dose that happen?
Hi Jianing,
Is puromycin and your gene of interest under one promoter? If you are using 2 promoters, it is possible that the promoter for your gene of interest is silenced.
Dear Jianing Liu
I hope these could be of relevance from previously raised Q on RG.
https://www.researchgate.net/post/Can_someone_advise_on_a_trouble_in_lentiviral_expression_GFP_expressed_target_gene_not
https://www.researchgate.net/post/Why_could_I_not_see_protein_overexpression_even_if_the_mRNA_increases
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