Hi everyone,
I am currently thinking of doing a genome wide CRISPR/Cas9 screen for drug resistance in cancer cells，but we have not done any research related to this in our lab. Although I have read some related papers( PMID: 24336571, PMID: 30414698, PMID: 28625976, ……), I still have a lot of problems. So, I urgently need your advice.
1. How to determine drug dose (such as, cisplatin) ?
2. How many days do I need to treat cancer cells with drug?
3. For negative selection, the drug dose and treatment time is the same as the positive selection?
4. Although there are protocols for genome wide CRISPR/Cas9 screen( PMID: 28333914 , PMID: 29224076), these protocols do not seem to explain these details. Are there other protocols?
If anyone can give me some advice, I will be very grateful. Thanks in advance!
1 You can take a gradient screening for it.for examples,the concentration that make the tumor in the edge of death and survival ,
2 Maybe you need some different tries
3 I think it should be same but it also has contact with your drug
4 you can use jove to find some protocols
I'm a rookie in library screen too,hope I can help you
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