Is it possible to express transgene in this lentiviral vector by cloning the cDNA after the EF1alpha promoter? The EF1a promoter which is driving the Mcherry expression and also the PuroR genes.
Yoav Lubelsky
Theoretically yes. Exactly how easy it will be depends on the exact vector sequence and whether or not you have convenient restriction sites. Also do you want to maintain the mCherry and/or Puro expression?
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Rishav Seal
Yes Yoav Lubelsky. I will be making a polycistronic expression with the MCherry and PuroR which will help me in sorting and then selecting the cells out.
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Yoav Lubelsky
How are the mCherry and Puro currently arranged? A fusion protein, 2A IRES?
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Yoav Lubelsky
In this case just remember to verify that all 3 genes are in the same reading frame with no stop codons between them.
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