I am planning to perform shRNA -mediated knockdown of my gene of interest using an inducible lentiviral based system. I am working on mouse embryonic stem cells. Can someone suggest a suitable vector for this model system? Has anyone used the pLKO Tet-On system? If so, can you suggest if I can clone a GFP sequence somewhere into this plasmid?
I have used retrovirus tet op.. check out the link http://www.ncbi.nlm.nih.gov/pubmed/24790089
Yes, I spent more than a year trying to do so! I HIGHLY recommend the pRSIT system from Cellecta. For a reasonable cost (from memory, it's $1200) you will get five vectors containing different shRNA sequences against your gene of interest. They are fully customisable with your choice of antibiotic resistance, GFP/RFP and dox-inducibility. Also you can choose your promoter. Best of all, even with all these features, the vectors are really small (8-9kb), which means that you can achieve higher lentiviral titers than if you package a large vector. Good luck! Let me know if you have any more questions.
Thank you for the responses. Ali, could you let me know the specific vector name that you have used for your purpose.
Hi Debrosee,
The vector backbone is pRSIT, and you can customise it to your liking. Speak to a representative from Cellecta and they can choose the perfect vector for you based on your needs.
Ali
Check the paper from the Elledge lab: http://www.ncbi.nlm.nih.gov/pubmed/21307310
They produced a set of pInducer vectors with shRNA cassettes under TRE promoter.
Cellecta is a great company that provides variety of services. The full list is provided on their website: https://www.cellecta.com/products-services/ Feel free to contact them and talk about best options for you and your experiments. They are friendly and the quality is the best one on the market.
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