CaMKK beta CRISPR Plasmids (h)human CaMKKβ-specific CRISPR/Cas9 KO Plasmid, HDR Plasmid, Double Nickase Plasmids, CRISPR Activation Plasmids and Lentiviral Activation Particles

CaMKKβ CRISPR Plasmids (h)

CaMKKβ CRISPR Knockout and Activation Products (h) is rated 5.0 out of 5 by 1.
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Datasheets

    • Target species: human; for corresponding mouse product, see CaMKKβ CRISPR Plasmids (m)
    • CRISPR/Cas9 KO Plasmids consists of CaMKKβ-specific 20 nt guide RNA sequences derived from the GeCKO (v2) library
    • For CRISPR gene knockout, gRNA sequences direct the Cas9 protein to induce a site-specific double strand break (DSB) in the genomic DNA
    • Target-specific CRISPR Plasmids for both gene knockout and activation are available. Please refer to the detailed product information in the tabs below
    • Gene knockdown or activation can be assayed using CaMKKβ Antibody (C-11): sc-271674
    • All products are provided as transfection-ready, purified plasmid DNA, except the Lentiviral Activation Particles which have been prepackaged as Lentiviral Particles for use with hard-to-transfect cells
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Rated 5 out of 5 by from A fabulous CRSPR based CamKK-beta knockout system. I am just so excited knocking out a gene using CRISPR/Cas9 technology for the first time in my research career that I thought I must share this experience. I used the SCBT CRISPR/Cas9 constructs (sc-400928) targeted to CaMKK-beta in human. These constructs are a mixture of three guide-RNAs (see attached figure A) targeted to ex-4 and ex-13 and can be effective to all isoforms. In addition, I have used the HDR plasmids (sc-400928-HDR) which has puro and RFP for positive selection (see fig A). The transfection was super easy and highly efficient in my choice of human carcinoma cell line, and the expression of RFP (HDR construct) and GFP (CRISPR construct) was perfect. Next, I selected 30-50 clones within 1-2 weeks of culture using puromycin and screened for CaMKK-beta expression using anti-CaMKK-beta specific antibody (sc-100364) from SCBT. Interestingly some of the selected clones expressed RFP and some not, but both RFP and non-RFP expressing cells were effectively knocked out for all isoforms (see fig B, RFP expressing clones marked red). The top 2 bands are CaMKK-beta isoforms and the bands disappeared in experiment in which I have used siRNA targeted to CaMKK-beta to knockdown the gene (see fig C). The CaMKK-beta antibody (sc-100364) is highly selective and does not cross react with homologous CaMKK-alpha protein (see fig C middle panel). The top 2 bands in fig B/C represent two isoforms of CaMKK-beta, and the bottom band is non-specific (ns), probably from secondary antibody. The bottom band disappeared in isoelectric focusing/ 2D SDS-PAGE followed by Immunoblot using CaMKK-beta specific antibody (sc-100364) see fig D. SO, IN SUMMARY, THE SANTA CRUZ CRISPR/CAS9 SYSTEM IS AWESOME AND IT DOES WHAT IT SUPPOSED TO. I HAVE FULL CONFIDENCE IN THEIR DESIGN AND WILL USE THEIR CRISPR PRODUCTS IN FUTURE TO KNOCKOUT OTHER GENES. Hope this review will help to make up your mind. Enjoy and cheers.
Date published: 2017-04-04
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