Cas9 Protein with NLS
CRISPR Cas9 Gene Editing
Cas9 protein, in complex with guide RNA, offers benefit for CRISPR/Cas9 technology over plasmid or mRNA transfection. The biggest advantage is that by incubating Cas9 protein and sgRNA briefly, you can form an RNP complex that is ready to work in vivo as soon as it is delivered into the cells or an embryo. Cas9 protein is short lived (about 24 hours of half-life in cells), thus minimizing the off target effects and mosaicism. You can also easily test the activity of guide RNA in vitro using our Cas9 protein by a simple cleavage assay. In addition, Cas9 protein is pretty stable, and activity confirmation is much easier compared to mRNA. Lastly, it is active in a more broad range of organisms such as nematodes, insects, plants, and non-typical model systems as it does not require transcription or translation.
Cas9 protein offered by PNA Bio is conjugated with NLS, which allows efficient transport of CRIRPR/Cas9 complex inside of the nucleus. Our Cas9 nuclease is provided as an injection/transfection ready form.
Cy3-Cas9 can be a useful tool for an easy enrichment of the cells that are transfected with CRISPR/Cas9 protein complex.
For in vitro cleavage protocol using PNA Bio's Cas9 protein, please download the following file. 
In vitro cleavage assay
PCR product was incubated with various amounts of Cas9 protein and run on the gel. In vitro cleavage assay is a simple way to check the activity of sgRNA.
Transfection of RNP complex
RNP complex of sgRNA and various amounts of Cas9 protein was transfected into NIH3T3 cells by electroporation (Neon) or by lipofectamine (LF2000).
Injection of RNP complex
For mouse embryo injection, RNP complex of 60 ng/ul of Cas9 protein and 15 ng/ul each of two sgRNAswas injected into a mouse embryo. The tail DNA was prepped from the pups and subject to T7E1 assay.Grey represents WT, green for large deletion (#2,3,4,8), red for small in/del (#5,6,7), blue for homozygous small in/del (#9, 10).
"We Guarantee the Activity and the Lowest Pricing of Cas9 Protein"
Cas9 Proteins
- Cas9 protein from Streptococcus pyogenes with NLS
- Provided as a lyophilized form
- Injection, transfection ready
- Activity validated by in vitro cleavage assay, electroporation (RNP complex), and mouse embryo injection
- Standard concentration (CP01): reconstitutes to 1 mg/ml, available as 20 ug, 50 ug, or 200 ug (50 ug x 4 tubes)
- High concentration (CP02 & CP03): reconstitutes to 5 mg/ml, available as 250 ug (CP02) or 1 mg (CP03, 250 ug x 4)
- Supplied with a Nuclease free water with 20% glycerol for reconstitution. Please request if you prefer Nuclease free water (for microinjection)
- CP01 is recommended for microinjection; CP02 or CP03 is preferred for electroporation
- Cas9 protein products are usually shipped the same day for the next day delivery
- For larger quantities, please contact us at order@pnabio.com or click here.
- Download in vitro cleavage protocol using PNA Bio's Cas9 protein
Cas9 Protein Buffers
- CB01: 10x Cas9 Buffer for in vitro cleavage assay, 500 ul each vial
- CB03: Cas9 Dilution Buffer when lower concentration of Cas9 protein is desired
- Cas9 protein buffers are usually shipped the same day for the next day delivery
Liquid form of Cas9 protein
- Available as 1 mg/ml, or 5 mg/ml
- CP04-100: 1 mg/ml (6 uM), 20 ug x 5 tubes
- CP04-500: 5 mg/ml (30 uM), 100 ug x 5 tubes
- Liquid form of Cas9 proteins are usually shipped the same day for the next day delivery
Nickase and dCas9 protein
- D10A Cas9 Nickase protein with NLS from Streptococcus pyogenes (CN01 & CN02)
- D10A/H840A Cas9 cleavage dead protein with NLS from Streptococcus pyogenes (CD01 & CD02)
- Available as 50 ug (reconstitutes to 1 mg/ml) or 250 (reconstitutes to 5 mg/ml) ug
- Nickase and dCas9 proteins are usually shipped the same day for the next day delivery
- For larger quantities, please contact us at order@pnabio.com or click here.
High Fidelity SniperCas9 protein
- Cas9 protein variant with high fidelity without compromising on activity
- Discovered by directed evolution method
- Available as 1 mg/ml (CSP01), or 5 mg/ml (CSP02)
- Read recent publication about SniperCas9
Cy3 labeled Cas9 protein
- Cy3 labeled Cas9 protein with NLS to enable sorting and visualization of transfected cells (CP06)
- Fluorescence detected by confocal microscopy
- Activity confirmed by in vitro cleavage assay and cell transfection
- Available as 50 ug, 100 ug or 250 ug sizes
- Concentration at 5 mg/ml
- Supplied with a Dilution buffer in case lower concentration is desired
- Please request a quote to order@pnabio.com or click here.
The activity of Cy3-Cas9 protein was confirmed by electroporation into cells followed by T7E1 assay. Fluorescence was confirmed by confocal microscopy.
| Product | Description | Available Sizes |
| CP01 | Cas9 protein with NLS (injection ready); reconstitutes to 1 mg/ml (6 uM) |
20 ug (120 pmol / 6 uM x 20 ul) 50 ug (300 pmol / 6 uM x 50 ul) 200 ug (50 ug x 4 tubes) |
| CP02 High conc Cas9 protein |
Cas9 protein with NLS (injection ready); reconstitutes to 5 mg/ml (30 uM) |
250 ug (1530 pmol / 30 uM x 50 ul) |
| CP03 High conc Cas9 protein |
Cas9 protein with NLS (injection ready); reconstitutes to 5 mg/ml (30 uM) |
1 mg (250 ug x 4) |
| CP04-100 Cas9 protein |
Cas9 protein with NLS (injection ready); Liquid form, 1 mg/ml (6 uM) |
100 ug (20 ug x 5) |
| CP04-500 High conc Cas9 protein |
Cas9 protein with NLS (injection ready); Liquid form, 5 mg/ml (30 uM) |
500 ug (100 ug x 5) |
| CS01 SniperCas9 protein |
HF Cas9 protein with NLS (injection ready); reconstitutes to 1 mg/ml (6 uM) |
50 ug (300 pmol / 6 uM x 50 ul) 200 ug (50 ug x 4 tubes) |
| CS02 High conc SniperCas9 protein |
HF Cas9 protein with NLS (injection ready); reconstitutes to 5 mg/ml (30 uM) |
250 ug (1530 pmol / 30 uM x 50 ul) |
| CN01 nickase protein |
Cas9 D10A Nickase protein with NLS (injection ready); reconstitutes to 1 mg/ml (6 uM) | 50 ug (300 pmol) |
| CN02 nickase protein |
Cas9 D10A Nickase protein with NLS (injection ready); reconstitutes to 5 mg/ml (30 uM) |
250 ug (1530 pmol) |
| CD01 dCas9 protein |
Cas9 D10A/H840A protein with NLS (injection ready); reconstitutes to 1 mg/ml (6 uM) |
50 ug (300 pmol) |
| CD02 dCas9 protein |
Cas9 D10A/H840A protein with NLS (injection ready); reconstitutes to 5 mg/ml (30 uM) |
250 ug (1530 pmol) |
| CP06 Cy3-Cas9 protein |
Cy3 labeled Cas9 protein with NLS (injection ready); 5 mg/ml solution (30 uM) |
50 ug/ 100 ug / 250 ug |
| CB01 Cas9 reaction buffer |
10X Cas9 cleavage reaction buffer | 500 ul |
| CB03 Cas9 dilution buffer |
Cas9 dilution buffer | 1 ml |
Publications referring our Cas9 protein |
- Peptide Nucleic Acid-Mediated Regulation of CRISPR-Cas9 Specificity. Carufe KEW et al. (2024) Nucleic Acid Ther In press.
- DNMT1 mutant ants develop normally but have disrupted oogenesis. Ivasyk I et al. (2023) Nat Commun 14(1): 2201.
- Spinal cord repair is modulated by the neurogenic factor Hb-egf under direction of a regeneration-associated enhancer. Cigliola V et al. (2023) Nat Commun 14:4857.
- CRISPR/Cas9 mediated genome editing of Caenorhabditis nigoni using the conserved dpy-10 co-conversion marker. Choi CP & Villeneuve AM (2023) span style="font-style:italic;font-weight:bold"> microPublication Biology 10:17912.
- CRISPR-enhanced human adipocyte browning as cell therapy for metabolic disease. Tsagkaraki E et al. (2021) Nat Commun 26; 12(1): 6931.
- Regulatory gene function handoff allows essential gene loss in mosquitoes. Cheatle AM et al. (2020) Commun Biol 3(1): 540.
- CRISPR/Cas9-mediated knock-in of alligator cathelicidin gene in a non-coding region of channel catfish genome. Simora RMC et al. (2020) Sci Rep 10(1): 22271.
- Simple embryo injection of long single-stranded donor templates with the CRISPR/Cas9 system leads to homology-directed repair in Xenopus tropicalis and Xenopus laevis. Nakayama T et al. (2020) Genesis 58(6): e23366.
- Multiplex CRISPR/Cas screen in regenerating haploid limbs of chimeric Axolotls. Sanor LD et al (2020) eLife 9: e48511.
- Germline CRISPR/Cas9-mediated gene editing prevents vision loss in a novel mouse model of aniridia. Mohanna SZM et al (2020) Mol Ther Methods Clin Dev 17: P478-490.
- Non-viral delivery of CRISPR/Cas9 complex using CRISPR-GPS nanocomplexes. Jain PK et al (2019) Nanoscale 11:21317-21323.
- Programmable CRISPR-Cas repression, activation, and computation with sequence-independent targets and triggers. Jin M et al. (2019) ACS Synthetic Biology 8(7):1583-1589.
- CRISPR/Cas9 gene editing in the West Nile Virus vector, Culex quinquefasciatus Say. Anderson ME at al. (2019) pLoS One 0224857.
- CRISPR/Cas9 Genome Editing Introduction and Optimization in the Non-model Insect Pyrrhocoris apterus. Kotwica-Rolinska J et al. (2019) Front Physiol 10: 00891.
- Small molecule agonists of Ae. aegypti neuropeptide Y receptor block mosquito biting. Duvall LB et al. (2019) Cell 176(4): 687.
- CRISPR/Cas9 F0 Screening of Congenital Heart Disease Genes in Xenopus tropicalis. Engin D et al. (2018) Methods in Molecular Biology 1865: 163.
- Microinjection of CRISPR/Cas9 Protein into Channel Catfish, Ictalurus punctatus, Embryos for Gene Editing. Elaswad A et al. (2018) J Vis Exp (131): e56275.
- Tissue-specific gene inactivation in xenopus laevis: knockout of lhx1 in the kidney with CRISPR/Cas9. DeLay BD et al. (2018) Genetics 208(2):673-686.
- A Non-integrating lentiviral approach overcomes cas9-induced immune rejection to establish an immunocompetent netastatic renal cancer model. Hu J et al. (2018) Mol Ther Methods Clin Dev 23(9):203-210.
- High prevalence of Streptococcus pyogenes Cas9-reactive T cells within the adult human population. Wagner DL et al. (2018) Nature Medicine 25:242–248.
- Application and optimization of CRISPR–Cas9-mediated genome engineering in axolotl (Ambystoma mexicanum) Fei J et al. (2018) Nat Protocols 13: 2908–2943.
- One‐step CRISPR/Cas9 method for the rapid generation of human antibody heavy chain knock‐in mice. Lin YC et al. (2018) 37:e99243 EMBO J 37:e99243.
- CRISPR knockouts reveal an endogenous role for ancient neuropeptides in regulating developmental timing in a sea anemone. Nakanishi N & Martindale MQ (2018)eLife 7: e39742.
- Antagonistic BMP–cWNT signaling in the cnidarian Nematostella vectensis reveals insight into the evolution of mesoderm. Wijesena N et al. (2017) Proc Natl Acad Sci USA 114: E5608–E5615.
- Generation of heritable germline mutations in the jewel wasp Nasonia vitripennis using CRISPR/Cas9. Li M et al. (2017) Scientific Reports 7:901.
- Generation of biallelic F0 mutants in medaka using the CRISPR/Cas9 system. Sawamura R et al. (2017) Genes to Cells 22:756–763.
- Laminar flow downregulates Notch activity to promote lymphatic sprouting. Choi D et al. (2017) J Clin Invest 127(4):1225-1240.
- Localized TWIST1 and TWIST2 basic domain substitutions cause four distinct human diseases that can be modeled in C. elegans. Kim S et al. (2017)
Hum Mol Genet 26(11): 2118–2132. - Genetic Basis of Melanin Pigmentation in Butterfly Wings. Zhang L et al. (2017) Genetics205(4):1537-1550.
- Efficient CRISPR/Cas9-assisted gene targeting enables rapid and precise genetic manipulation of mammalian neural stem cells. Bressan RB et al. (2017) Development 144(4):635-648.
- CRISPR/Cas9-mediated gene editing in human zygotes using Cas9 protein. Tang L et al. (2017) Mol Genetics Genomics 292(3):525-533.
- A truncating mutation in CEP55 is the likely cause of MARCH, a novel syndrome affecting neuronal mitosis. Frosk P et al. (2017) J Med Genet 54(7):490-501.
- Quantitative Analysis of Synthetic Cell Lineage Tracing Using Nuclease Barcoding. Schmidt ST et al. (2017) ACS Synth Biol 6(6):936–942.
- Targeted gene knock-in by CRISPR/Cas ribonucleoproteins in porcine zygotes. Park KE et al. (2017) Hum Mol Genet 7:42458.
- Maternal Supply of Cas9 to Zygotes Facilitates the Efficient Generation of Site-Specific Mutant Mouse Models. Cebrian-Serrano A et al. (2017) PLoS One0169887.
- Molecular logic behind the three-way stochastic choices that expand butterfly colour vision. Perry M et al. (2016) Nature 535(7611):280-4.
- Leapfrogging: primordial germ cell transplantation permits recovery of CRISPR/Cas9-induced mutations in essential genes. Blitz IL et al. (2016) Development 143(15):2868-75.
- Delivery of Cas9 Protein into Mouse Zygotes through a Series of Electroporation Dramatically Increases the Efficiency of Model Creation. Wang W et al. (2016) J Genetics Genomics 43(5)319-327.
- Efficient genome engineering approaches for the short-lived African turquoise killifish. Harel I et al. (2016) Nature Protocols. 11(10):2010-28.
- Rapid Screening for CRISPR-Directed Editing of the Drosophila Genome Using white Co-Conversion. Ge DT et al. (2016) G3 (Bethesda) 6(10): 3197–3206.
- A truncating mutation in CEP55 is the likely cause of MARCH, a novel syndrome affecting neuronal mitosis. Frosk P et al. (2016) J Medical Genetics 104296.
- Generation and characterization of tamoxifen-inducible Pax9-creER knock-in mice using CrispR/Cas9. Jifan F et al. (2016) Genesis 54: 490–496.
- Expanding the genetic toolkit in Xenopus: Approaches and opportunities for human disease modeling. Tandon P et al. (2016) Dev Biology S0012-1606.
- Dual daughter strand incision is processive and increases the efficiency of DNA mismatch repair. Hermans N et al. (2016) Nucleic Acids Res 44(14):6770-86.
- Genome editing in butterflies reveals that spalt promotes and Distal-less represses eyespot colour patterns. Zhang L & Reed RD (2016) Nat Commun 7:11769.
- Pre-bilaterian origin of the blastoporal axial organizer. Kraus Y et al. (2016) Nat Commun 7:11694.
- CRISPRs for optimal targeting: delivery of CRISPR components as DNA, RNA, and protein into cultured cells and single-cell embryos. Kouranova E et al. (2016) Hum Gene Ther 27(6):464-75.
- Highly efficient genome editing of murine and human hematopoietic progenitor cells by CRISPR/Cas9. Gundry MC et al. (2016) Cell Reports 17(5):1453-61.
- Genomic Access to Monarch Migration Using TALEN and CRISPR/Cas9-Mediated Targeted Mutagenesis. Markert MJ et al. (2016)G3 (Bethesda) 6(4):905-15.
- A CRISPR Path to Engineering New Genetic Mouse Models for Cardiovascular Research. Miano JM et al. (2016) Arterioscler Thromb Vasc Biol 36(6):1058-75.
- CRISPR/Cas9-mediated mutagenesis in the sea lamprey Petromyzon marinus: a powerful tool for understanding ancestral gene functions in vertebrates. Square T et al. (2015) Development 142(23):4180-7.
- High Efficiency, Homology-Directed Genome Editing in Caenorhabditis elegans Using CRISPR/Cas9 Ribonucleoprotein Complexes. Paix A et al. (2015) Genetics 201(1):47-54.
- CRISPR-CAS9 D10A nickase target-specific fluorescent labeling of double strand DNA for whole genome mapping and structural variation analysis. McCaffrey J et al. (2015) Nucleic Acids Res 44(2):e11.
- Cell lineage tracing using nuclease barcoding. Schmidt ST et al. (2016) Cornell University Library Epub.
- Silencing of end-joining repair for efficient site-specific gene insertion after TALEN/CRISPR mutagenesis in Aedes aegypti. Basu S et al. (2015) Proc Natl Acad Sci U S A 112(13):4038-43.
- In vivo Modeling Implicates APOL1 in Nephropathy: Evidence for Dominant Negative Effects and Epistasis under Anemic Stress. Anderson BR et al. (2015) PLoS Genet 11(7):e1005349.
- CRISPR/Cas9: An inexpensive, efficient loss of function tool to screen human disease genes in Xenopus. Bhattacharya D et al. (2015) Dev Biology 408(2): 196-204.
- Genome engineering with CRISPR-Cas9 in the mosquito Aedes aegypti. Kistler KE et al. (2015) Cell Reports 11(1): 51-60.
- Adenovirus-Mediated Somatic Genome Editing of Pten by CRISPR/Cas9 in Mouse Liver in Spite of Cas9-Specific Immune Responses. Dan W et al. (2015) Hum Gene Ther 26(7): 432-442.
- Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells. Hendel A et al. (2015) Nat Biotech 33:985-89.
- Cloning-free CRISPR/Cas system facilitates functional cassette knock-in in mice. Aida T et al. (2015) Genome Biology 16:87.
- TALEN and CRISPR/Cas9-mediated genome editing in the early-branching metazoan Nematostella vectensis. Ikmi A et al. (2014) Nat Commun 24(5):5486.
- Chapter Seventeen: Cas9-Based Genome Editing in Xenopus tropicalis. Nakayama T et al. (2014) Methods Enzymol (Editor J A Doudna & E. J. Sontheimer) 546:355-375.
Books referring our Cas9 protein |
- Genome Editing & Engineering: From TALENs, ZFNs and CRISPRs to Molecular Surgery (ed. Appasani K) Leapfrogging: Targeting CRISPR/Cas9 Genome Editing to the Germline in Xenopus. Blitz IL (2018) Cambridge University Press
- Diversity and Evolution of Butterfly Wing Patterns (ed. Sekimura T & Nijhout HF) A Practical Guide to CRISPR/Cas9 Genome Editing in Lepidoptera. Zhang L & Reed RD (2017) pp 155-172. Springer
- The Cricket as a Model Organism; Development, Regeneration, and Behavior by Horch, H.W., Mito, T., Popadić, A., Ohuchi, H., Noji, S. (2017) Springer
- Methods in Cell Biology, The Zebrafish: Genetics, Genomics, and Transcriptomics, Volume 135, 4th Edition (Detrich HW, Zon L. & Westerfield M) (2016) Academic Press
On-line manuals using our Cas9 protein |
- Practical guide to genome-engineering with CRISPR-Cas9 in the mosquito Aedes aegypti (Kistler KE et al.) - Rockefeller University
- KI mice generation - Beth Israel Deaconess Medical Center Transgenic Core Facility
- RNA guide prep and purification - University of Wisconsin-Madison Biotechnology Center Transgenic Animal Facility
- Optimization of DNA, RNA and RNP Delivery for Efficient Mammalian Cell Engineering - Mirus Bio
- CRISPR-based gene disruptionin human hematopoietic cells - The Goodell Laboratory
Cat No
Item
Description
Size
Price
Order
CP01-200
Cas9 protein with NLS
Reconstitutes to 1 mg/ml,
1200 pmol (6 uM x 200 ul)
200 ug (50 ug x 4)
$325.00
CP02
Cas9 protein with NLS, high concentration
Reconstitutes to 5 mg/ml,
1530 pmol (30 uM x 50 ul)
250 ug
$340.00
CP03
Cas9 protein with NLS, high concentration
Reconstitutes to 5 mg/ml,
6000 pmol (30 uM x 50 ul x 4 tubes)
1 mg (250 ug x 4)
$1,150.00
CP04-100
Cas9 protein with NLS, Liquid form, 100 ug (20 ug x 5)
Liquid form 1 mg/ml, 600 pmol
(6 uM x 20 ul x 5 tubes)
100 ug (20 ug x 5)
$220.00
CP04-500
Cas9 protein with NLS, Liquid form, 500 ug (100 ug x 5)
Liquid form 5 mg/ml, 3000 pmol
(30 uM x 20 ul x 5 tubes)
500 ug (100 ug x 5)
$695.00
CS01-200
SniperCas9 (HF)
High Fidelity Cas9 protein,
Reconstitutes to 1 mg/ml
200 ug (50 ug x 4)
$385.00
CN01
D10A Cas9 nickase protein with NLS
Reconstitutes to 1 mg/ml,
300 pmol (6 uM x 50 ul)
50 ug
$115.00
CN02
D10A Nickase protein with NLS, high concentration
Reconstitutes to 5 mg/ml,
1530 pmol (30 uM x 50 ul)
250 ug
$350.00
CD01
Dead Cas9 (D10A/H840A) protein with NLS
Reconstitutes to 1 mg/ml,
300 pmol (6 uM x 50 ul)
50 ug
$115.00
CD02
Dead Cas9 (D10A/H840A) protein with NLS, high concentration
Reconstitutes to 5 mg/ml (30 uM), 1530 pmol (30 uM x 50 ul)
250 ug (50 ug x 5)
$350.00