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  1. Theoretically speaking, can you insert the gene sequences for cas9, sgRNA, and promoters into the host genome so that the cell perpetually produces CRISPR components?

  2. In this scenario, I'm guessing there would be no way to have the host cell produce DNA templates for HDR?

  3. How long do CRISPR plasmids last in the nucleus and is there any way have them persist for 10+ days?

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    $\begingroup$ Quick answers/comments: 1) yes 2) why not? 3) stable transfection/genomic insertion using retro or lentiviruses. If the insertion is stable, it can last the life of the organism. $\endgroup$
    – MattDMo
    Commented Jan 6, 2023 at 17:29
  • $\begingroup$ Comment to 1: Yes. Many labs have integrated Cas9 stabely into their cell lines. E.g., for CRISPR/Cas9 screens (forward genetic screens). It's very useful, since then you can avoid double transfections (of Cas9 and sgRNA) which lowers the efficiency. $\endgroup$
    – Felix H.
    Commented Aug 23, 2023 at 7:58

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