Plasmid pCas9(D10A) from Dr. Xiao Wang's lab contains the insert Cas9 D10A and is published in ACS Synth Biol. 2015 Nov 20;4(11):1217-25. doi: 10.1021/acssynbio.5b00132. Epub 2015 Oct 25. This plasmid is available through Addgene.

The CRISPR Cas9-D10A Nickase expression plasmids use the CMV promoter for strong transient expression of Cas9. Alternate promoters can be substituted by replacement of CMV using MluI and NheI. Also, the Cas9-D10A expression plasmids can be linearized using XbaI for T7-based mRNA production.

Feb 15, 2015 · Plasmid hCas9_D10A from Dr. George Church's lab contains the insert Cas9_D10A and is published in Science. 2013 Feb 15;339(6121):823-6. doi: 10.1126/science.1232033 This plasmid is available through Addgene.

Plasmid pDe-CAS9-D10A from Dr. Holger Puchta's lab contains the insert Cas9-D10A and is published in Plant J. 2014 Jul;79(2):348-59. doi: 10.1111/tpj.12554. Epub 2014 Jun 17. This plasmid is available through Addgene.

Mammalian vector with a G418 resistance marker for expressing human codon-optimized Cas9 with the D10A nickase mutation.

Plasmid for mammalian cell expression of Cas9 with the D10A nickase mutation and with an N-terminal nuclear localization signal followed by a FLAG® tag.

The Cas9-D10A nickase plasmid co-expressing GFP uses the CMV promoter for strong transient expression of Cas9-D10A. Alternate promoters can be substituted by replacement of CMV using MluI and NheI. Also, the Cas9D10A expression plasmids can be linearized using XbaI for T7-based mRNA production.

Plasmid for reconstituting the S. pyogenes CRISPR/Cas system in E. coli. Use together with pCRISPR.

Recently, the type II CRISPR/Cas system from the bacterium Streptococcus pyogenes has been engineered to function in eukaryotic systems using two molecular components: a single Cas9 protein and a non-coding guide RNA (gRNA).The Cas9 endonuclease can be rendered inactive (dCas9) with mutations to the two protein domains, RuvC and HnH (D10A and ...

Mar 15, 2018 · Overview of Cas9 nickase. We’ll use SpCas9 nickases as examples for this post. The D10A mutation inactivates the RuvC domain, so this nickase cleaves only the target strand. Conversely, the H840A mutation in the HNH domain creates a non-target strand-cleaving nickase.