A recent revolution in biology has been stimulated by the discovery of CRISPR, or “Clustered Regularly Interspaced Short Palindromic Repeats” and the understanding of the “CRISPR Associated” enzymes (CAS 1,2).
The CRISPR repeated sequences are found in bacterial genomes and function as part of unique bacterial immune system which contain short DNA sequences derived from viruses which have infected the bacteria.
These virally derived sequences can make short RNA sequences which can hybridize with specific viral DNA and target a nuclease, such as CAS9, to the viral sequence.
So CAS9 is directed to cleave the specific viral sequence and so inactivate the virus.
The RNA sequence can be designed to specifically cut DNA virtually anywhere, including in the genomes of living human and other mammalian cells, allowing inexpensive gene editing with unprecedented ease.
For example three groups of researchers essentially cured the disease state in a mouse model of Duchenne muscular dystrophy (3-5).
A similar approach essentially cured dogs affected with a related disease state (6).
Several varieties of CAS9 have been studied and there are several other related enzymes with similar properties.
Much of the early work was performed with CAS9 from Streptococcus pyogenes (7).
The S.
pyogenes protein is rather large at 1,368 amino acids, ~158kDa, so the corresponding DNA is also rather large at about 4.2kb.
This is problematic with some expression systems especially since DNA encoding RNA sequences and possibly other regulatory elements are usually required.
The CAS9 gene of Staphylococcus aureus is significantly smaller, at about 3kb, producing a protein of 124kDa (7).
For an excellent recent review of the various CAS family enzymes and their utility see reference 8.
Applications:Suitable for use in Western Blot, Immunohistochemistry, Immunofluorescence and Immunocytochemistry.
Other applications not tested.
Recommended Dilution:Immunofluorescence/Immunocytochemistry: 1:1000-1:5000Western Blot: 1:1000Optimal dilutions to be determined by the researcher.
Storage and Stability:May be stored at 4℃ for short-term only.
Aliquot to avoid repeated freezing and thawing.
Store at -20℃.
Aliquots are stable for 12 months after receipt.
For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
仕様
Size：100ul
抗体種類：Mab
宿主：mouse
Grade：Affinity Purified
Purity：Purified by Protein G affinity chromatography.
Form：Supplied as a liquid in PBS, 0.03% sodium azide, 50% glycerol.
Specificity：Recognizes Cas9 in Staphylococcus aureus. Does not recognize Cas9 in Streptococcus pyogenes.
Clone # USB：6F7
Isotype：IgG1
Calc Applications Abbrev：IC IF IHC WB
Immunogen：Recombinant protein fragment corresponding to 251aa, aa803-1053, from the C-terminal region of Staphylococcus aureus Cas9, expressed in E. coli.
