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FNCAS9PROT

Sigma-Aldrich

FnCas9 Protein

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

Synonym(s):
Francisella Novicida Cas9

Quality Level

recombinant

expressed in E. coli

assay

≥95% (SDS-PAGE)

form

lyophilized powder

packaging

pkg of 1 kit (4 components)

application(s)

CRISPR

shipped in

wet ice

storage temp.

−20°C

Related Categories

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. 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

Legal Information

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

Kit Components Only

Product No.
Description

  • FnCas9-NLS from Francisella novicida, expressed in Escherichia coli

  • Dilution buffer for Cas9 proteins

  • Reconstitution solution for Cas9 proteins

  • Digestion buffer 10× for FnCas9

Pictograms

Exclamation mark

Signal Word

Warning

Hazard Statements

Hazard Classifications

Eye Irrit. 2

Storage Class Code

11 - Combustible Solids

WGK

WGK 3

Certificate of Analysis

Enter Lot Number to search for Certificate of Analysis (COA).

Certificate of Origin

Enter Lot Number to search for Certificate of Origin (COO).

Quotes and Ordering

Protocols

Ribonucleoprotein (RNP) Protocols Using Synthetic sgRNAs and Cas9 proteins

Combine guaranteed sgRNAs with our comprehensive range of CRISPR products and tools, including Cas9 and delivery reagents, for efficient genome modification with higher specificity.

Related Content

Visualize your CRISPR Experiment with MISSION

Watch Cas9-GFP proteins work inside your cells and deliver the powerful nuclease editing you have come to trust and expect. MISSION™ SpCas9-GFP and eSpCas9-GFP proteins are multi-functional enzymes with high on-target cleavage efficiency fused to Enhanced Green Fluorescent Protein (EGFP) for real-time observation of Cas9 delivery and clearance from cells.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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