Need help: electroporation to transform CRISPR plasmids (sgRNA, CAS9 and HDR) doesn't work (algae)

I met some problems with CRISPR-CAS9 system for microalgae.

I've read lots of papers about this technology. But I can hardly find papers using electroporation to transform CRISPR vectors into algae. Instead, most of the papers used biolistic bombardments.

Electroporation is more frequently used for RNP(ribonucleoprotein complexes) transformation. Why? I tried electroporation to transform vectors carrying sgRNA, CAS9 and HDR and I got few colonies! Anyone has experience in it? Electroporation doesn't work for CRISPR vector system??

Thank you!


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Mint Wyvernover 3 years ago

Electroporation will work with the CRISPR vector system.

The RNP has it's own benefits such as decreased CAS exposure (less availability to get off target due to less time CAS hangs around), no need to "cure" a plasmid, and the RNP CAS seems to be highly specific. If you have the ability to use electroporation many people are choosing that method for those reasons. If electroporation is not an option then there is nothing inherently wrong with using another transformation method.

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