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문서

FNCAS9P

Sigma-Aldrich

FnCas9 plasmid

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About This Item

UNSPSC 코드:
41106609
NACRES:
NA.51

재조합

expressed in E. coli

포장

vial of 50 μL

농도

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

응용 분야

CRISPR

프로모터

Promoter name: CMV

selection

kanamycin

배송 상태

dry ice

저장 온도

−20°C

일반 설명

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

애플리케이션

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

특징 및 장점

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

원리

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.

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 2

Flash Point (°F)

Not applicable

Flash Point (°C)

Not applicable


시험 성적서(COA)

제품의 로트/배치 번호를 입력하여 시험 성적서(COA)을 검색하십시오. 로트 및 배치 번호는 제품 라벨에 있는 ‘로트’ 또는 ‘배치’라는 용어 뒤에서 찾을 수 있습니다.

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문서 라이브러리에서 최근에 구매한 제품에 대한 문서를 찾아보세요.

문서 라이브러리 방문

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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