Cas9 Protein with NLS

Cas9 protein, in complex with guide RNA, offers benefit for CRISPR/Cas9 technology over plasmid or mRNA transfection. The biggest benefit is that you can make RNP complex that is ready to work in vivo. Cas9 protein is short lived (about 24 hours of half life in cells), thus minimize the off-target effects and mosaicism. You can also easily test the activity of guide RNA in vitro 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 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, and it is injection/transfection ready form.

Cy3-Cas9 can be an easy enrichment of the cells that are transfected with CRISPR/Cas9 complex.

In vitro cleavage assay

Cas9 protein NLS cleavage assay with sgRNA

PCR product was incubated with various amounts of Cas9 protein and run on the gel. In vitro cleavageassay is a simple way to check the activity of sgRNA.

Transfection of RNP complex

Cas9 protein NLS cleavage assay with sgRNA

RNP complex of sgRNA and various amount of Cas9 protein was transfected into NIH3T3 cells by electropration (Neon) or by lipofectamine (LF2000).

Injection of RNP complex

Cas9 protein NLS cleavage assay with sgRNA

For mouse embryo injection, RNP complex of 60 ng/ul of Cas9 protein and 15 ng/ul each of two sgRNAswas injected into 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 forhomozygous 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
  • Injection, transfection ready
  • Activity validated by in vitro cleavage assay, electroporation (as RNP complex), and mouse embryo
    injection for each batch
  • Standard concentration (CP01, reconstitutes to 1 mg/ml): 20 ug, 50 ug, or 200 ug (50 ug x 4)
  • High concentration (CP02 & CP03, reconstitutes to 5 mg/ml): 250 ug (CP02) or 1 mg (CP03, 250 ug x 4)
  • Packaging: 20 ug or 50 ug for CP01 and 250 ug for CP02/CP03
  • CP01 is recommended for microinjection and CP02/CP03 is recommended for electroporation
  • Also available: D10A Nickase (CN01) and D10A/H840A dCas9 (CD01) protein
  • Cas9 protein is usually shipped the same day if order is received by 5 pm EST
  • For larger quantities, please contact us at order@pnabio.com or click here.


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 size, reconstitutes to 1 mg/ml (CN01 / CD01) or 250 ug size, reconstitutes to 5 mg/ml (CN02 / CD02).
  • For larger quantities, please contact us at order@pnabio.com or click here.


Cy3-Cas9 & Cy3-dCas9 protein

  • Cy3 labeled Cas9 protein with NLS (CP06) to enable sorting and visualization of transfected cells
  • Fluorescence detected by confocal microscopy
  • Activity confirmed by in vitro cleavage assay and cell transfection
  • Available as 50 ug (CP06-50) or 200 ug (CP06-200) at 5 mg/ml concentration
  • Also Cy3 labeled dCas9 protein (CD07) is available
  • Please request a quote to order@pnabio.com or click here.

Cy3-Cas9 protein NLS cleavage assay with sgRNA

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
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)
CN01
nickase protein
Cas9 D10A Nickase protein injection ready with NLS; reconstitutes to 1 mg/ml (6 uM) 50 ug
CN02
nickase protein
Cas9 D10A Nickase protein injection ready with NLS; reconstitutes to 5 mg/ml (30 uM) 250 ug
CD01
dCas9 protein
Cas9 D10A/H840A protein with NLS;
injection ready, reconstitutes to 1 mg/ml (6 uM)
50 ug
CD02
dCas9 protein
Cas9 D10A/H840A protein with NLS;
injection ready, reconstitutes to 5 mg/ml (30 uM)
250 ug
CP06
Cy3-Cas9 protein
Cy3 labeled Cas9 protein with NLS;
5 mg/ml solution (30 uM)
50 ug
200 ug
CD07
Cy3-dCas9 protein
Cy3 labeled Cas9 dead protein with NLS;
5 mg/ml solution (30 uM)
50 ug
200 ug

Publications referring our Cas9 protein

  1. Laminar flow downregulates Notch activity to promote lymphatic sprouting. Choi D et al. (2017)  J Clin Invest In press.
  2. 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 In press.
  3. Genetic Basis of Melanin Pigmentation in Butterfly Wings. Zhang L et al. (2017) Genetics In press.
  4. Efficient CRISPR/Cas9-assisted gene targeting enables rapid and precise genetic manipulation of mammalian neural stem cells. Bressan RB et al (2017) Development In press.
  5. CRISPR/Cas9-mediated gene editing in human zygotes using Cas9 protein. Tang L et al (2017) Mol Genetics Genomics In press.
  6. 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 In press.
  7. Quantitative Analysis of Synthetic Cell Lineage Tracing Using Nuclease Barcoding. Schmidt ST et al. (2017) ACS Synth Biol In press.
  8. Targeted gene knock-in by CRISPR/Cas ribonucleoproteins in porcine zygotes. Park KE et al. (2017) Hum Mol Genet 7:42458.
  9. Maternal Supply of Cas9 to Zygotes Facilitates the Efficient Generation of Site-Specific Mutant Mouse Models. Cebrian-Serrano A et al (2017) PLoS One 0169887.
  10. Molecular logic behind the three-way stochastic choices that expand butterfly colour vision. Perry M et al (2016) Nature 535(7611):280-4.
  11. 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 In press.
  12. Efficient genome engineering approaches for the short-lived African turquoise killifish. Harel I et al. (2016) Nature Protocols. 11(10):2010-28.
  13. 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.
  14. 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.
  15. Generation and characterization of tamoxifen-inducible Pax9-creER knock-in mice using CrispR/Cas9. Jifan F et al. (2016) Genesis 54: 490–496.
  16. Expanding the genetic toolkit in Xenopus: Approaches and opportunities for human disease modeling. Tandon P et al. (2016) Dev Biology S0012-1606.
  17. 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.
  18. Genome editing in butterflies reveals that spalt promotes and Distal-less represses eyespot colour patterns. Zhang L & Reed RD (2016) Nat Commun 7:11769.
  19. Pre-bilaterian origin of the blastoporal axial organizer. Kraus Y et al. (2016) Nat Commun 7:11694.
  20. 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.
  21. 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.
  22. Genomic Access to Monarch Migration Using TALEN and CRISPR/Cas9-Mediated Targeted Mutagenesis. Markert MJ et al. (2016)G3 (Bethesda) 6(4):905-15.
  23. 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.
  24. 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.
  25. High Efficiency, Homology-Directed Genome Editing in Caenorhabditis elegans Using CRISPR/Cas9 Ribonucleoprotein Complexes. Paix A et al. (2015) Genetics 201(1):47-54.
  26. 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.
  27. Cell lineage tracing using nuclease barcoding. Schmidt ST et al. (2016) Cornell University Library Epub.
  28. 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.
  29. 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.
  30. 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.
  31. Genome engineering with CRISPR-Cas9 in the mosquito Aedes aegypti. Kistler KE et al. (2015) Cell Reports 11(1): 51-60.
  32. 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.
  33. Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells. Hendel A et al. (2015) Nat Biotech 33:985-89.
  34. Cloning-free CRISPR/Cas system facilitates functional cassette knock-in in mice. Aida T et al. (2015) Genome Biology 16:87.
  35. TALEN and CRISPR/Cas9-mediated genome editing in the early-branching metazoan Nematostella vectensis. Ikmi A et al. (2014) Nat Commun 24(5):5486.
  36. 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

1. Methods in Cell Biology, The Zebrafish: Genetics, Genomics, and Transcriptomics, Volume 135, 4th Edition by Detrich HW, Zon L. & Westerfield M. (2016) Academic Press
2. The Cricket as a Model Organism; Development, Regeneration, and Behavior by Horch, H.W., Mito, T., Popadić, A., Ohuchi, H., Noji, S. (2017)

On-line manuals using our Cas9 protein

1.    Practical guide to genome-engineering with CRISPR-Cas9 in the mosquito Aedes aegypti (Kistler KE et al.) - Rockefeller University
2.    KI mice generation - Beth Israel Deaconess Medical Center Transgenic Core Facility
3.    RNA guide prep and purification - University of Wisconsin-Madison Biotechnology Center Transgenic Animal Facility
4.    Optimization of DNA, RNA and RNP Delivery for Efficient Mammalian Cell Engineering (Khodthong C et al.)
5.    CRISPR/Cas9 editing: cotransfection of Cas9 protein and synthetic RNA oligos
Cat No
Item
Description
Size
Price
Order
CP01-20
Cas9 protein with NLS

Reconstitutes to 1 mg/ml,

120 pmol (6 uM x 20 ul)

20 ug
$75.00
CP01-50
Cas9 protein with NLS

Reconstitutes to 1 mg/ml,

300 pmol (6 uM x 50 ul)

50 ug
$115.00
CP01-200
Cas9 protein with NLS

Reconstitutes to 1 mg/ml,

1200 pmol (6 uM x 200 ul)

200 ug (50 ug x 4)
$350.00
CP02
Cas9 protein with NLS, high concentration

Reconstitutes to 5 mg/ml,

1530 pmol (30 uM x 50 ul)

250 ug
$400.00
CP03
Cas9 protein with NLS, high concentration

Reconstitutes to 5 mg/ml, 

6000 pmol (30 uM x 200 ul)

1 mg (250 ug x 4)
$1,200.00
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 high concentration

Reconstitutes to 5 mg/ml,

1530 pmol (30 uM x 50 ul)

250 ug
$400.00
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, high concentration

Reconstitutes to 5 mg/ml, 

1530 pmol (30 uM x 50 ul)

250 ug
$400.00
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