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詳細
The Cas9 expression plasmids use the CMV promoter for strong transient expression of Cas9. Alternate promoters can be substituted by replacement of CMV using MluI and NheI. Also, the Cas9 expression plasmids can be linearized using XbaI for T7-based mRNA production. The addition of a fluorophore that is translationally co-expressed with the Cas9 nuclease allows for easy visualization of successful transfection.
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
構成
1 vial containing 1 μg of Cas9-2A-RFP plasmid.
Please note, product does not contain guideRNA sequence. This must be purchased separately through the Custom CRISPR product tab.
Please note, product does not contain guideRNA sequence. This must be purchased separately through the Custom CRISPR product tab.
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.
物理的形状
Sigma Cas9-2A-RFP plasmid DNA is supplied at concentrations of 50ng/μl in 20μl.
その他情報
Must be used in conjunction with a U6-gRNA plasmid in order to mediate a double strand break in the DNA.
Typical transfection concentrations used in literature are in the ranges of >= 1.0 μg/μL and <= 5 μL of Cas9 plasmid combined with >= 1.0 μg/μL and <= 5 μL of U6-gRNA plasmids. (All dosages above assume 0.5 to 1 million cells nucleofected).
Typical transfection concentrations used in literature are in the ranges of >= 1.0 μg/μL and <= 5 μL of Cas9 plasmid combined with >= 1.0 μg/μL and <= 5 μL of U6-gRNA plasmids. (All dosages above assume 0.5 to 1 million cells nucleofected).
保管分類コード
10 - Combustible liquids
WGK
nwg
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
CAS9RFPP-1EA:
CAS9RFPP-BULK:
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試験成績書（COA）
Lot/Batch Number
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プロトコル
Learn about CRISPR Cas9, what it is and how it works. CRISPR is a new, affordable genome editing tool enabling access to genome editing for all.
関連コンテンツ
Sigma-Aldrich® Advanced Genomics is the leading provider of gene editing and silencing technologies including CRISPR, Cas9, synthetic guide RNA (sgRNA), and Zinc Finger Nuclease (ZFN).
Sigma-Aldrich®アドバンストゲノミクスは、CRISPR、Cas9、合成ガイドRNA（sgRNA）、およびジンクフィンガーヌクレアーゼ（ZFN）などの遺伝子編集とサイレンシング工学技術の有力なプロバイダーです。
アクティブなフィルタ
ライフサイエンス、有機合成、材料科学、クロマトグラフィー、分析など、あらゆる分野の研究に経験のあるメンバーがおります。.
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