FnCas9 plasmid FNCAS9P-1UG

This product is an expression plasmid that utilizes the CMV promoter for strong transient expression of Francisella novicida Cas9 (CMV-FnCas9). The FnCas9 expression plasmid is one part of a two part CRISPR system with individual FnCas9 and gRNA expression vectors.
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Functional Genomics/Target Validation

• Proxy CRISPR
• Creation of gene knockouts in multiple cell lines

Features and Benefits

• Highly specific and highly active
• Sequence verified
• Ready to use purified plasmid DNA

Principle
CRISPR/Cas systems are employed by bacteria and archaea as a defense against invading viruses and plasmids. Recently, the type II-B CRISPR/Cas system from the bacterium Francisella novicida has been engineered to function in eukaryotic systems using two molecular components: a single FnCas9 nuclease and a non-coding guide RNA (gRNA). The FnCas9 endonuclease is human codon optimized and programmed with a gRNA, directing a DNA double-strand break (DSB) at a desired sequence of DNA. Similar to DSBs induced by zinc finger nucleases (ZFNs), the cell then activates endogenous DNA repair processes, either non-homologous end joining (NHEJ) or homology-directed repair (HDR), to heal the targeted DSB. FnCas9 is similar to common nucleases in many ways but also possess other distinct qualities. It has been determined to possess a higher intrinsic specificity through a reduced tolerance to gRNA:DNA mismatches as well as inducing a staggered cleavage pattern leaving 4bp 5′ overhangs by cleaving the non-target strand at 7 bp from the PAM. In contrast to SpCas9, FnCas9 activity fluctuates across genomic regions. Cotargeting dead Cas9 at proximal locations may alter local chromatin structures and render otherwise inaccessible target sites accessible and cleavable by FnCas9.
CRISPR Label License,

recombinant	expressed in E. coli
packaging	vial of 50 µL
concentration	20 ng/µL in TE buffer; DNA (1µg of plasmid DNA)
application(s)	CRISPR: suitable
Promoter	Promoter name: CMV
selection	kanamycin
shipped in	dry ice
storage temp.	20°C

RIDADR	NONH for all modes of transport
WGK Germany	WGK 2
Flash Point F	Not applicable
Flash Point C	Not applicable