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リコンビナント
expressed in E. coli
包装
vial of 50 μL
濃度
20 ng/μL in TE buffer; DNA (1μg of plasmid DNA)
アプリケーション
CRISPR
プロモーター
Promoter name: CMV
セレクション
kanamycin
輸送温度
dry ice
保管温度
−20°C
詳細
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
To order gRNA in any format click here
アプリケーション
Functional Genomics/Target Validation
- Proxy CRISPR
- Creation of gene knockouts in multiple cell lines
特徴および利点
- Highly specific and highly active
- Sequence verified
- Ready to use purified plasmid DNA
原理
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.
保管分類コード
12 - Non Combustible Liquids
WGK
WGK 2
引火点(°F)
Not applicable
引火点(℃)
Not applicable
適用法令
試験研究用途を考慮した関連法令を主に挙げております。化学物質以外については、一部の情報のみ提供しています。 製品を安全かつ合法的に使用することは、使用者の義務です。最新情報により修正される場合があります。WEBの反映には時間を要することがあるため、適宜SDSをご参照ください。
Jan Code
FNCAS9P-1UG:
FNCAS9P-BULK:
最新バージョンのいずれかを選択してください：
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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