I am new to the CRISPR Cas9 genome editing system and I have the above basic question.

Another way to think about this: what would happen if after cells are transfected, the cultures are left to grow without any selection. Thus, if a mutation disrupts a cell cycle gene (e.g. p53), such cells will proliferate more and the culutre will be populated by those cells rather than healthy cells.

Is this a valid reasoning?

  • 2
    $\begingroup$ Your suggestion of using CRISPR to disrupt P53 and hope for selecting cells that grow out of control isn't what most CRISPR experiments aim to do. CRISPR is used to get precise mutations, but doesn't always work. Only a small percent of cells will be successfully edited, so you have to screen for them and find a good one. This is what makes CRISPR so hard to use in vivo, where you can't just kill off 99.99% of cells and let the good ones grow back. $\endgroup$ – user137 May 12 '16 at 15:27
  • $\begingroup$ user137 is correct. Screening for successful CRISPR/CAS9 mutation is a headache. You typically need to either titrate your cells down to 1 cell/well of a 96-well plate, let them expand and genotype, or something more rigorous like this: bmcgenomics.biomedcentral.com/articles/10.1186/… $\endgroup$ – akaDrHouse May 12 '16 at 15:35

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