Why doesn't transfection with CRISPR/Cas show any GFP expression?

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Mahmudul Hasan

If I understand correctly from your question, is your sgRNA fused to an mRNA expressing GFP? If that is true, my hypothesis is when you co-transfect, The cas9 forms a ribonuclear complex with the gRNA itself, making it unavailable for GFP expression.

I would recommend a fluorophore tagged gRNA instead of using mRNA. Or may be use a dCas9 mRNA fused to GFP using P2A. Hope this helps.

Phan Thi Lam Hong

Thank you for your help~