I would like to perform the in vitro synthetic lethality screen in cancer cells to identify the synthetic lethal pair. Is there any HTS method for identification of SL pair? and also any method to determine the anticancer compound that inhibiting the codependent gene or protein of SL pair?
MTT assay would work. https://www.promega.de/products/cell-health-and-metabolism/cell-viability-assays/celltiter-96-non_radioactive-cell-proliferation-assay-_mtt_/
I used this kit for cytotoxicity assay, and it worked very well
Dear Dr. Andrey Luchnik,
I would be pleased to hear the answer from you.
Best regards,
Hi Thokhir Basha Shaik,
Q: How do I perform an in vitro synthetic lethality screening for cancer cells?
A: We conducted a genome wide siRNA screen - see below publication
Q: and also any method to determine the anticancer compound that inhibiting the codependent gene or protein of SL pair?
A: We conducted a 4,057 known drug screen and time course studies on a CDH1 isogenic MCF10A cell lines - see below publication
Hope this helps,
Henry Beetham
Article Synthetic Lethal Screens Identify Vulnerabilities in GPCR Si...
It is established that the dysfunction of Na+/K+-pump is a common consequence of any pathology, including cancer. At the same time, it is also known that among a number of mechanisms involved in cell volume regulation, Na+/K+-pump has a central role in it. This role of pump is due to its two important properties: a) Na+/K+-pump which generates Na+ gradient on the membrane serves as an energy source for a number of secondary ionic transporters, such as, Na+/Ca2+, Na+/H+, Na+/sugars, amino acids & other osmolytes and b) Na+/K+-pump, having the highest metabolic energy (ATP) utilizing mechanism, has a great intracellular signaling role in regulation of sorption properties of intracellular structure as well as in generation of water molecules during oxidative glucose processes in cells. Na+/K+-ATPase (working molecules of Na+/K+-pump) has 4 catalytic subunits having different functional activities and sensitivities to cardiac glycoside: α1 (low), α2 (middle), α3 and α4 (high), the latter is identified only in testis. From these isoforms α1 (fully) and α2 (partly) have ion transporting functions, while α3 isoform only performs intracellular signaling function. By previous our study it was shown that dysfunction of α3 isoform-dependent signaling function controlling cell hydration stimulates carcinogenesis: So if you can measure the level of cell hydration it can be indicator for cancer screening
Thank you very much to all for their answers it helped me a lot.
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