Hi Community,
I am planning to express 6 human proteins in a mouse cell line. They are from 400 to 800 bp long. I want to use three vectors, each containing two CDS connecting with 2A peptides, followed by IRES promoter controlling various fluorescent proteins. However, I am just wondering if having three of them would affect the efficiency of the processing of 2A peptides and causes a severe downregulation of these genes. Transducing 6 genes separately means I have to go through many more clones to get one that expresses all due to lack of selection.
Has anyone have any experience on multiple multicistronic gene expression?
Hi Yannick Frey ,
Thanks for the clarification! I always thought it was like a peptide because the Liu and colleagues article and others used self-cleaving. Always read the references I guess!
Can I ask with your experience, does the size of the gene matter? i.e. are smaller genes less prone to the loss of expression level in the second position compared to their longer counterpart?
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