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Key Documents

FNCAS9P

Sigma-Aldrich

FnCas9 plasmid

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About This Item

分類程式碼代碼:
41106609
NACRES:
NA.51

重組細胞

expressed in E. coli

包裝

vial of 50 μL

濃度

20 ng/μL in TE buffer; DNA (1μg of plasmid DNA)

應用

CRISPR

啟動子

Promoter name: CMV

選擇

kanamycin

運輸包裝

dry ice

儲存溫度

−20°C

一般說明

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.

To order gRNA in any format click here

應用

Functional Genomics/Target Validation
  • Proxy CRISPR
  • Creation of gene knockouts in multiple cell lines

特點和優勢

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

原則

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.

儲存類別代碼

12 - Non Combustible Liquids

水污染物質分類(WGK)

WGK 2

閃點(°F)

Not applicable

閃點(°C)

Not applicable


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Structure and Engineering of Francisella novicida Cas9
Hirano, H. et al.
Cell, 164, 950-961 (2016)
Targeted activation of diverse CRISPR-Cas systems for mammalian genome editing via proximal CRISPR targeting.
Chen, F. et al.
Nature Communications, 8 (2017)

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