I would like to insert a 1500 bp or longer gene to the broken site in E.coli (or may be bacillus subtilis which support NHEJ) after cut with Crispr-Cas9. Is this possible for both NHEJ or HDR?

  • $\begingroup$ CRIPSR-Cas9 can be used in eukaryotes to insert much larger fragments (using two different sgRNAs and HDR), so in in principle this should be possible - I'm not sure if it will work in bacteria though. $\endgroup$ – Nicolai Nov 24 '18 at 14:21
  • $\begingroup$ @Nicolai thank you very much, can only one sgRNA work? $\endgroup$ – joe Nov 24 '18 at 15:20

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