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
Schematic Diagram of Cas9-sgRNA and Target DNA Complex
NLS-Cas9-NLS
Cas9(D10A)-Nickase
-Nickase.jpg "Cas9(D10A)-Nickase")
dCas9
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
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