CRISPR vitro system - (Mar/10/2014 )

Hey all,

Does any know if there is a company or a group that is selling or able to provide the CRISPR-CAS9 protein?

I would like to use the CRISPR system invitro to cut DNA. I can't use restriction enzymes.

Thanks

here are a few companies that seem to have what you want:

sigma

origene

system biosciences

(maybe) mirus bio

Thank Mdfenko, but not quite what I was looking for.What I really want is the CAS9 protein itself. Like how one obtains and uses an enzyme. like Taq polymerase for PCR.

I'm  look for somebody who is actually selling the protein, I want to avoid to time and technicality required to express and purify the CAS9 protein.

Hi,I have a quick question.Can Cas9 protein cut DNA without guideRNA?

Chen Wang on Mon Mar 24 09:59:19 2014 said:

Hi,I have a quick question.Can Cas9 protein cut DNA without guideRNA?

I'm not certain on this, but I don't believe it can. I work with a dCas9 protein and it doesn't seem to bind to anything in my sgRNA-free controls. It would be pretty a pretty lousy immune system in bacteria if it did, too. Perhaps you could create a Cas9 enzyme with a sgRNA fused to it somehow? But that's beyond my depth.

That's another challenge in using a straight up Cas9 protein to cut DNA too, I think, since I'm not sure you'll be able to imitate the secondary structure of the sgRNA either. 

perneseblue on Mon Mar 10 19:48:35 2014 said:

Hey all,

 

Does any know if there is a company or a group that is selling or able to provide the CRISPR-CAS9 protein?

I would like to use the CRISPR system invitro to cut DNA. I can't use restriction enzymes.

 

Thanks

That is a very interesting idea. I am not sure whether earlier papers studying CRISPR have tried this like RNAi people have for in vitro RNAi. In this in vitro system, you need to add guide RNA which could be chemically synthesized.

Edit: Just found a paper: In vitro assembly and activity of an archaeal CRISPR-Cas type I-A Cascade interference complex.