Is it possible to generate stable isolated clones of SH-SY5Y Ko cells line (with CRISPR/Cas9) using FACS (mCherry) ?

11 November 2016 1 5K Report

We want to make stable SH-Sy5y Cell line Ko cell line after Transfection with CRISPR/Cas9. We looking the best way to isolate our transfected clones (FACS or Puromycin).

Tom Masi

Does your FACs machine have a green/yellow laser in it? If not, you will not be able to "see' the mCherry signal. The standard blue laser only hits the shoulder of the excitation peak of mCherry and the signal is weak/non-existent. Puromycin selection is probably a better option if you do not have the correct laser for optimal mCherry excitation.

Good Luck!

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