Description
General description
The purified FnCas9 protein (~190 kDa) is derived from the type II CRISPR/Cas system from the bacterium Francisella novicida and has been engineered to function using three molecular components: the FnCas9 protein, a crRNA of user-defined sequence, and a tracrRNA of constant sequence. The FnCas9 endonuclease can be programmed by changing the crRNA sequence to create a site-specific double-strand break (DSB) in plasmids or purified genomic DNA.
Application
Functional Genomics/Target Validation/Genome Editing/Molecular Biology/Molecular Cloning/Synthetic Biology
Biochem/physiol Actions
Conventional restriction enzymes such as EcoRI and HindIII can only cut specific short (6 bp) DNA sequences and are therefore limited to cloning small DNA fragments (typically ~4 kb). CRISPR-based endonculeases can cut user defined sequences of a longer length (~20 bp) making them more flexible and applicable to larger DNA fragments such as multigene bacterial operons or mammalian genomic DNA with native promoter, exon, and intron components.
Features and Benefits
Suitable for plasmid or large genomic DNA fragment cloning
Results in 4-bp 5′ overhangs.
Cuts purified plasmid DNA more efficiently than SpCas9
Packaging
pkg of 50 μg (≥ 300 pmol)
pkg of 250 μg (≥ 1500 pmol)
Preparation Note
Lyophilized FnCas9 protein should be resuspended in the Reconstitution solution provided to desired concentration. Gently tap tube to completely dissolve lyophilized powder, incubate for 10 minutes on ice, and spin tube to bring material to bottom of tube.
Other Notes
Click here to order gRNA
Enter the 20bp genomic DNA target sequence (5′ to 3′) plus the adjacent 3′ PAM site. Under special instructions, indicate "FnCas9 crRNA"
Click here to order FnCas9 tracrRNA
Check out our other MISSION® Cas9 Proteins at SigmaAldrich.com/CRISPRproteins
Each kit consists of:
one vial of FnCas9 recombinant protein
one vial containing 1 mL of 1× Dilution buffer
one vial containing 1 mL of Nuclease free water with glycerol
one vial containing 1 mL of 10× Digestion Buffer
Legal Information
CRISPR Use License Agreement
MISSION is a registered trademark of Merck KGaA, Darmstadt, Germany
