pX330 vs pX459

[DG]

Hi all,

is there any difference in efficiency of transfection and/or protein synthesis/effect of the Cas9 between pX330 and pX459? Or do they work exactly the same?

Thanks and best

[Le Cong]

Hi, they are based on the same design and if you compare seqs, the Cas9 cassette is almost identical (promoter, epitope tags), except the additional 2A-Puro part, and the U6 part are also basically exact match so one would expect most parameters to be similar. One note is 2a linker will leave the Cas9 part to have an additional ~20aa peptide attached to it, which usually won't affect protein activity much.

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Le CONG, Ph.D. 丛乐

Broad Institute of MIT and Harvard
HHMI International Fellow

Tel: (+1) 617-823-6132
E-Mail: congl...@gmail.com / co...@broadinstitute.org

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[DG]

Hi Le,

correct, everything is the same but for the puromycin, which is pretty small. So in theory you could expect the same effect

But have you (or anyone) indeed compare them in any experiment?. Is there the same success rate?