The plasmid is for lentivirus, higher protein expression should be constitutive. I have the plasmid with wild-type gene. I know there is another way to increase gene expression constitutively, for example VP16 fusion gene. My question is based on the wild-type target gene, how to use mutagenesis to receive up-regulated expression, not via VP16 or other fusion gene. Any commercial kit is available for this aim?
Hello see below info may be useful for you.
https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0014408
Good luck!
Do you consider changing promoter? We have created a more flexible lentivirus vector that you can do modifications easily. Please contact noahgen.com if you are interested.
Hello see below info may be useful for you.
https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0014408
Good luck!
Hi Jianxuan,
I wonder if you have solved the problem you encountered in the experiment.
I can recommend you a company that can do mutagenesis, and they have many types of services. https://www.creative-biostructure.com/custom-mutagenesis-services-341.htm
You can also ask if they have the appropriate commercial kit. I feel very good, you can have a try.
Good luck!
Karen A. Darbinyan Thanks for your answer with PLOS paper. That paper may not meet my needs. I am using lentivirus carrying wild-type gene or wild-type plus up/down regulated gene to stably infect myeloma and see the effects from different expression level. In that PLOS paper, they found PC3 could achieve higher expression than 293T, and could be down-regulated by some inhibitors.
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