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FNCAS9PROT Sigma-Aldrich

FnCas9 Protein

from Francisella novicida, recombinant, expressed in E. coli, 1X NLS

Synonym: Francisella Novicida Cas9

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Properties

Related Categories CRISPR-Cas9, Functional Genomics and RNAi, MISSION™ Cas9 Proteins, Molecular Biology
Quality Level   200
recombinant   expressed in E. coli
assay   ≥95% (SDS-PAGE)
form   lyophilized powder
application(s)   CRISPR: suitable
shipped in   wet ice
storage temp.   −20°C

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

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)

Components

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

Principle

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.

Reconstitution

Lyophilized FnCas9 protein should be resuspended in the Reconstitution solution provided to desired concentration. The solution should be allowed to incubate at room temperature (20-23 °C) for 10 minutes. Centrifuge immediately before use.

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

Legal Information

CRISPR Use License Agreement

MISSION is a registered trademark of Sigma-Aldrich Co. LLC

Kit component only

Description

Product #

Add to Cart

FnCas9-NLS from Francisella novicida, expressed in Escherichia coli    
Dilution buffer for Cas9 proteins    
Reconstitution solution for Cas9 proteins    
Digestion buffer 10× for FnCas9    
Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
Protocols & Articles

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