What are the differences in genome-wide CRISPR library vs specified CRISPR library?

I am trying to make custom CRISPR library to find specifically transcription factors. I am wondering if I don't need to make another library for transcription factors rather than genome-wide targets.

Because I think the genome-wide library make more background gRNA abundance than a custom library which is minimized and specified.

How do you think about that? Do you think I need to do screening with the genome-wide library?

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Olive Cerberusover 3 years ago

Briefly, can I increase the possibility to find the target?

Olive Fenrirover 3 years ago

Theoretically you can, because you can set higher coverage.

I think, more important thing is it can reduce your labor and experimental misses.

Cells grow very rapidly than I expected.

Fuchsia Kelpieover 3 years ago

If you only care about transcription factors, then it would make sense to screen only sgRNAs targeting transcription factors, because this will reduce your experimental cost and labor. It will also reduce the "noise" in your screen, and can potentially reduce your false positive rate.

We provide a python script for isolating sgRNAs targeting the genes of interest from an existing sgRNA library, and experimental protocol for cloning the custom library in Joung Nat Protocol 2017 that you might find helpful for your work.

Olive Cerberusover 3 years ago

I am thinking about what is the best way to find the target that I want to know. I should design a new library precisely.

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