Im researching the role of gene TRIM58 in enucleation process in hematophoeisis. I did already a knock-out of TRIM58 but i would like to do a cell line where i overexpress it too. What would be the best way to do so? CRISPR coupled with a transcription factor? Any other suggestions? Thanks for the help
One of the common ways of overexpressing a gene is to clone it into an expression construct and transfect and retain the plasmid in the cell using a selectable marker. You can find a compatible episomal expression vector by reading up on papers that have done so on the cell line you are working on.
Hi Marko Klimez ,
Cloning viral transcription factor factor, for example, 'tat' gene (Trans-Activator of Transcription) of HIV in front of your target gene can help overexpression of some proteins. I worked with some tat-transduced cell lines that exhibit overexpression.
Please see below the workflow for theTRIM58 Stable Cell Line Generation
-Host cell line: You may choose any (HEK293T or CHO-K1, or other cell types are also available according to your requirements)
-Vector construction
-Pool cells generation
-Monoclonal cells screening and identification
-Validation by qPCR
-Mycoplasma detection
Have fun and Happy research!
- Badges
- Science topic
- Similar topics
- Physical Sciences
- Materials Science
- Materials
- Carbon Materials
- Graphite
- Graphene