NLS-dCas9-EGFP-NLS

NLS-dCas9-EGFP-NLS
PR-137213G
$245.00

$350.00

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Size
50 ug
Storage
-80°C or -20 °C Avoid freeze thaw cycles.
Application
In situ hybridization, subcellular localization of defined sequence, enrich cell populations for desired genome edits via EGFP-based fluorescence activated cell sorting (FACS).
Background
NLS-dCas9-EGFP is a fusion protein containing a nuclear localization sequence (NLS) on its N terminal and EGFP on the C terminal. The dCas9 RNP complex can localize to the nucleus immediately upon entering the cell with the addition of a nuclear localization signal (NLS). The EGFP can be taken as a reporter for tracking or sorting transfected cells, which creates the possibility of enriching cell populations for desired genome edits via fluorescence activated cell sorting (FACS). NLS-dCas9-EGFP is better than NLS-Cas9-EGFP in genome tracking by in situ hybridization, due to the inactivity of nuclease while retaining the DNA binding ability
Purity
Greater than 90% as determined by SDS-PAGE
Host/Source
E.coli
Concentration
1 µg/µL
storage buffer
10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM DTT, 300 mM NaCl, and 50% (v/v) Glycerol


1.  Deactivated CRISPR Associated Protein 9 for Minor-Allele Enrichment in Cell-Free DNA    Amin Aalipour et al. Clinical Chemistry 2018 Vol. 64, Issue 2 p307-p916
2. Purified Cas9 Fusion Proteins for Advanced Genome Manipulation Jovan Mircetic et al. Small Methods 2017 1, 1600052
3. Disruptive non-disruptive applications of CRISPR/Cas9  Jonathan LSchmid-Burgk  Current Opinion in Biotechnology December 2017, Volume 48 Pages 203-209
4. Efficient sequence-specific isolation of DNA fragments and chromatin by in vitro enChIP  technology using recombinant CRISPR ribonucleoproteins Toshitsugu Fujita et al. Genes to Cells  Volume 21, Issue 4 April 2016  Pages 370–377
5. High-throughput biochemical profiling reveals sequence determinants of dCas9 off-target binding and unbinding Evan A Boyle  et al. PNAS 2017 May, 114 (21) 5461-5466
6.CASFISH: CRISPR/Cas9-mediated in situ labeling of genomic loci in fixed cells  Wulan Deng et al. PNAS 2015 vol. 112 | no. 38 p11870-11875

NLS-dCas9-EGFP-NLS

 

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