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Hello,I also want to purify AsCpf1, but I want to know whether...

Bahtiyar Kurtulmus Oct 16, 2018

Protocol: Heterologous Expression and Purification of the CRISPR-Cas12a/Cpf1 Protein

Hello,

I also want to purify AsCpf1, but I want to know whether you have tested a construct such as 6xHis-AsCpf1 for purification. Is MBP tag critical for the purification process (i.e. solubility). I do not want to perform TEV cleavage since I do not have a very good experience Read more more

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Daan C. Swarts Author Answered Jun 17, 2019

Department of Biochemistry, University of Zurich, Switzerland

Dear Bahtiyar,

I am curious if you were successful with the purification! Do you need any other input?

With kind regards,
Daan

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Daan C. Swarts Author Answered Oct 26, 2018

Department of Biochemistry, University of Zurich, Switzerland

Dear Bahtiyar,

Unfortunately, we have not tried purifying AsCas12a without MBP tag.

We have purified FnCas12a and other Cas12a variants with and without MBP tag, and did not see any significant changes in yield. Although this does not give any guarantees for AsCas12a, I think it is worth trying.

With kind regards,
Daan

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