Where should I design the gRNA if I want to insert point mutations with CRISPR/Cas9?

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Lahiru Chamara Weerasinghe Arachchige

You can try base editing technology or prime editing technology, rather than conventional DNA double-strand breaks. There are a lot of options available at the moment. It is faster and will have fewer off-targets. You can find more information from Dr David Liu's lab.

https://www.addgene.org/David_Liu/

For Prime editors (to design pegRNA)

http://pegfinder.sidichenlab.org/

For conventional methods, you can follow this webtool and their publication as reference.

http://crispor.tefor.net/

Romain Gioia

Hi. Thank you for your answer. Lahiru Chamara Weerasinghe Arachchige How many plasmid do I need to use the prime editing technology ? I don't find an all-in-one plasmid with pegRNA-Cas9-RT on your Addgene page.