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リコンビナント
expressed in E. coli
品質水準
包装
vial of 50 μL
濃度
20 ng/μL in TE buffer; DNA (1μg of plasmid DNA)
プロモーター
Promoter name: CMV
レポーター遺伝子
RFP
selection
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 eSpCas9 and RFP linked by a 2A peptide (CMV-eSpCas9-2A-RFP) allowing for easy visualization of successful transfection. The eSpCas9 expression plasmid is one part of a two part CRISPR system with individual eSpCas9 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
- Creation of gene knockouts in multiple cell lines
- Complete knockout of genes not amenable to RNAi
- Creation of knock-in cell lines with promoters, fusion tags, or reporters integrated into endogenous genes
特徴および利点
- Enhanced specificity compared to wild type Cas9
- Highly active
- 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 CRISPR/Cas system from the bacterium Streptococcus pyogenes has been engineered to function in eukaryotic systems using two molecular components: a single Cas9 protein and a non-coding guide RNA (gRNA). The Cas9 endonuclease can be programmed with a single gRNA, directing a DNA double-strand break (DSB) at a desired genomic location. 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. Newly engineered eSpCas9 enables the efficient targeted gene editing of established CRISPR systems with the benefit of reduced off-target effects. Point mutations in the chromosome-binding motif of SpCas9, as described by Slaymaker, et al., provide higher on-target fidelity without loss of cleavage efficiency.
保管分類コード
10 - Combustible liquids
WGK
WGK 2
引火点(°F)
Not applicable
引火点(℃)
Not applicable
試験成績書(COA)
製品のロット番号・バッチ番号を入力して、試験成績書(COA) を検索できます。ロット番号・バッチ番号は、製品ラベルに「Lot」または「Batch」に続いて記載されています。
Rationally engineered Cas9 nucleases with improved specificity.
Science, 351, 84-88 (2015)
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