I am trying to understand how CRISPR has made the gene knockout or gene editing process simpler to make transgenic animals. Here is an old (pre CRISPR) flowchart from Manis, 2007 that shows how knockouts can be made. enter image description here

I assume much of this process remains the same even with CRISPR. But I want to understand what part of this flowchart has become easier with CRISPR? Is it that we do not need to wait for a random recombination event to occur? Is it that the yield has increased because we directly cut at the site and ask for HDR to happen rather than just sit and wait for it to happen?

  • 2
    $\begingroup$ Before CRISPR, homologous recombination at the target site (first step in left chart) relied on such events happening spontaneously. With CRISPR, the ability to generate double-strand breaks (DSBs) at the target site dramatically increases the frequency of homologous recombination events, since the DSB activates the cell´s own DNA repair mechanisms (of which homologous recombination is one) and directs them to the target site. $\endgroup$
    – gaspanic
    Apr 23 at 23:40
  • 1
    $\begingroup$ The wiki page for gene knockout lists several non-CRISPR methods for gene knockouts including zinc-fingers. $\endgroup$
    – doremi
    Apr 24 at 21:22

Your Answer

By clicking “Post Your Answer”, you agree to our terms of service, privacy policy and cookie policy

Browse other questions tagged or ask your own question.