Characteristics of Streptococcus pyogenes Cas9 variants used for genome editing

Product name Catalog # CRISPR-Cas9 variants Amino acid features NLS position Application PAM sequence Amount Concentration Price  
NLS-Cas9-NLS PR-137211 Streptococcus pyogenes Cas9 wild type wild type N-terminal and C-terminal Create double blunt end for recombination NGG 50 µg, 313 pmol 1 µg/ µL, 6.26 µM $105 In stock
NLS-Cas9-EGFP PR-137211-E Streptococcus pyogenes Cas9 with C-terminal EGFP wild type N-terminal and C-terminal Create double blunt end for recombination NGG 50 µg, 265 pmol 1 µg/ µL, 5.31 µM $195 In stock
NLS-Cas9(D10A)-2NLS nickase PR-137212B Streptococcus pyogenes Cas9 nickase D10A N-terminal and 2xC-terminal Create nick on one strand NGG 50 µg, 313 pmol 1 µg/ µL, 6.26 µM $195 In stock
dCas9 (D10A & H840A) PR-137213 No nuclease activity while keeping target binding activity D10A/H840A C-terminal Detection and purification targeted DNA NGG 50 µg, 313 pmol 1 µg/ µL, 6.26 µM $250 In stock
NLS-dCas9-NLS PR-137213B No nuclease activity while keeping target binding activity D10A/H840A N-terminal and C-terminal Detection and purification targeted DNA NGG 50 µg, 313 pmol 1 µg/ µL, 6.26 µM $250 In stock
Biotinylated NLS-dCas9-NLS PR-137213-bio No nuclease activity while keeping target binding activity, biotinylated D10A/H840A N-terminal and C-terminal Specific DNA fragment pull-down in CHIP assay. FISH and short sequence dtection NGG 50 µg, 313 pmol 1 µg/ µL, 6.26 µM $250 In stock
dCas9-SBP PR-137213-SBP No nuclease activity while keeping target binding activity, biotinylated D10A/H840A C-terminal Specific DNA fragment pull-down in CHIP assay. FISH and short sequence dtection NGG 50 µg, 313 pmol 1 µg/ µL, 6.26 µM $250 In stock
NLS-dCas9-EGFP-NLS PR-137213G No nuclease activity while keeping target binding activity D10A/H840A N-terminal and C-terminal FISH, genomic visualization NGG 50 µg, 265 pmol 1 µg/ µL, 5.31 µM $245 In stock
Lb Cas12a (Cpf1) Nuclease PR-137215 Cut 5' TTTN PAM Sequence   N-terminal and C-terminal Cleavage 5' TTTN 50 µg 1 µg/ µL  $175 In stock
eSpCas9(1.1)-3NLS Cas9-HiFi1 Cas9 with increase On-target specificity K848A/K1003A/R1060A N-terminal and 2xC-terminal Gene editing with better on-target, less off-target rate. NGG 50 µg, 313 pmol 1 µg/ µL, 6.26 µM $195 2 business days
SpCas9-HF1-3NLS Cas9-HiFi2 Cas9 with increase On-target specificity N497A/R661A/Q695A/Q926A N-terminal and 2xC-terminal Gene editing with better on-target, less off-target rate. NGG 50 µg, 313 pmol 1 µg/ µL, 6.26 µM $195 2 business days
Cas9 protein CAS9-OEM           Any size   please inquire  
Cas9 protein ( choose one product above) Free Trial           10µg   Only pay Shiping and handling  

The Cas9 protein (CRISPR-associated protein 9), derived from type II CRISPR (clustered regularly interspaced short palindromic repeats) bacterial immune systems, is becoming an essential tool for engineering the genome in diverse organisms. As an RNA-guided DNA endonuclease, Cas9 is designated to target new sites by altering its guide RNA sequence, and its development as a tool has made sequence-specific gene editing much easier. The nuclease-deactivated form of dCas9 further provides a versatile RNA-guided DNA-targeting platform for regulating and imaging the genome in nucleus, as well as for rewriting the epigenetic dynamics, all in a sequence-specific manner. With all of these progress, scientists have begun to explore the possible applications of Cas9 in biomedical research and therapeutics. Novatein Biosciences is working hard to help you in fully utilize the Cas9 related proteins, antibodies, and assay kit to advance your projects. We focus on the applications of Cas9 proteins for genome editing, regulation, and subcellular localization of target genes.

 

Protocol: FISH or gene tracking in vitro with labeled dCas9, dCas9-GFP, dCas9-biotin

 

Protocol: Preparation of Cas9 RNP (ribonucleoprotein), transfection into adherent cells using the Lipofectamine

 

Schematic Diagram of Cas9-sgRNA and Target DNA Complex

NLS-Cas9-NLS

NLS-Cas9-NLS

Cas9(D10A)-Nickase

Cas9(D10A)-Nickase

dCas9

dCas9 target DNA binding activity assay

dCas9 target DNA binding activity assay


dCas9 competes with WT Cas9 for target binding

  • C. Linearized 7 kb plasmid DNA, 400 ng.
    • 1.100 ng WT Cas9 nuclease only
    • 2.500 ng dCas9 protein + 100 ng WT Cas9 nuclease
    • 3.250 ng dCas9 protein + 100 ng WT Cas9 nuclease
    • 4.125 ng dCas9 protein + 100 ng WT Cas9 nuclease
    • 5.62.5 ng dCas9 protein + 100 ng WT Cas9 nuclease
  • 30 ng of crRNA/tracrRNA mixed with various amount of dCas9 protein.
  • Then 400 ng substrate plasmid DNA was added and WT Cas9 protein followed.

NLS-dCas9-GFP-NLS

NLS-dCas9-GFP-NLS

A comprehensive Cas9 reagents ready to meet your research and development needs.
NLS-Cas9-NLS (Catalog number: PR-137211) transfection ready for gene editing, also used for sgRNA or crRNA screening (Catalog number: NB-GEN10642)

NLS-dCas9-NLS (Catalog number: PR-137213B) for precise gene regulation and interrogation.

NLS-Cas9-EGFP (Catalog number: PR-137211-E) for subcellular localization of the Cas9-gRNA/target DNA complex.

sgRNA Screening Kit (Catalog number: NB-GEN10642) for quickly assessing a sgRNA sequence in vitro to see if it can work effectively in vivo.

Anti-CRISPR/CAS9 Monoclonal Antibody (Catalog number: SH-A16960) detection of Cas9 ( wild type, nickase and dCas9) in WB, IF, and ELISA

Anti-CRISPR/CAS9 Polyclonal Antibody (Catalog number: SH-A16959) detection of Cas9 ( wild type, nickase and dCas9) in WB, IF, and ELISA

Cas9 ELISA kit (Catalog number: NB-E1372PR) to check if or how much Cas9 protein is transfected or expressed.

 

References

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?77

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


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