Cas9 (S. aureus) (6H4) Mouse mAb (Alexa Fluor® 647 Conjugate) #30004
- IF
Supporting Data
| REACTIVITY | All |
| SENSITIVITY | Transfected Only |
| MW (kDa) | |
| Source/Isotype | Mouse IgG2b |
Application Key:
- IF-Immunofluorescence
Species Cross-Reactivity Key:
- All-All Species Expected
Product Information
Product Description
Product Usage Information
| Application | Dilution |
|---|---|
| Immunofluorescence (Immunocytochemistry) | 1:50 |
Storage
Protocol
Specificity / Sensitivity
Cas9 (S. aureus) (6H4) Mouse mAb (Alexa Fluor® 647 Conjugate) recognizes endogenous levels of total Cas9 (S. aureus) protein. This antibody does not cross-react with Cas9 (S. pyogenes), AsCpf1 (Strain BV3L6), and FnCpf1 (Strain U112) proteins.
Species Reactivity:
All Species Expected
Source / Purification
Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of Cas9 (S. aureus) protein.
Background
The CRISPR associated protein 9 (Cas9) is an RNA-guided DNA nuclease and part of the CRISPR antiviral immunity system that provides adaptive immunity against extra chromosomal genetic material (1). The CRISPR antiviral mechanism of action involves three steps: (i), acquisition of foreign DNA by host bacterium; (ii), synthesis and maturation of CRISPR RNA (crRNA), followed by the formation of RNA-Cas nuclease protein complexes; and (iii), target interference through recognition of foreign DNA by the complex and its cleavage by Cas nuclease activity (2). The type II CRISPR/Cas antiviral immunity system provides a powerful tool for precise genome editing and has potential for specific gene regulation and therapeutic applications (3). The Cas9 protein and a guide RNA consisting of a fusion between a crRNA and a trans-activating crRNA (tracrRNA) must be introduced or expressed in a cell. A 20-nucleotide sequence at the 5' end of the guide RNA directs Cas9 to a specific DNA target site. As a result, Cas9 can be "programmed" to cut various DNA sites both in vitro and in cells and organisms. CRISPR/Cas9 genome editing tools have been used in many organisms, including mouse and human cells (4,5). Research studies demonstrate that CRISPR can be used to generate mutant alleles or reporter genes in rodents and primate embryonic stem cells (6-8).
Cas9 (S. aureus) is a Cas9 ortholog that is smaller, but as efficient, as the more commonly used Cas9 ortholog, Cas9 (S. pyogenes) (9).
- Horvath, P. and Barrangou, R. (2010) Science 327, 167-70.
- Wiedenheft, B. et al. (2012) Nature 482, 331-8.
- Singh, P. et al. (2015) Genetics 199, 1-15.
- Cong, L. et al. (2013) Science 339, 819-23.
- Mali, P. et al. (2013) Science 339, 823-6.
- Li, D. et al. (2013) Nat Biotechnol 31, 681-3.
- Shen, B. et al. (2013) Cell Res 23, 720-3.
- Niu, Y. et al. (2014) Cell 156, 836-43.
- Ran, F.A. et al. (2015) Nature 520, 186-91.
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