추천 제품
재조합
expressed in E. coli
Quality Level
분석
≥95% (SDS-PAGE)
형태
lyophilized powder
포장
pkg of 1 kit (3 components)
응용 분야
CRISPR
배송 상태
wet ice
저장 온도
−20°C
일반 설명
CAS9, also known as cas5 and csn1, is the signature gene of the type II clustered regularly interspaced short palindromic repeats (CRISPR)-RuvC (RNase H-like fold) cas system. CAS9 contains 1388 amino acids. This protein is predicted to contain a RuvC/ ribonuclease (RNase) H domain involved in crRNA maturation and McrA/HNH signature domain involved in the DNA degradation step.
Recombinant Cas9 protein from Streptococcus pyogenes (~160 KD) is a ready-to-use reagent for genome engineering experiments. When combined with target-specific guide RNAs, wild type Streptococcus pyogenes Cas9 protein will act as a targeted nuclease suitable for transfection of cell cultures and for the accelerated development of genetically-modified animals via one-cell embryo injection.
애플리케이션
Functional Genomics/Target Validation/Genome Editing
생화학적/생리학적 작용
CAS9 plays a vital role in plasmid DNA interference. It is the only cas protein needed to deliver resistance against foreign DNA. CAS9 stimulates both RNA-guided genome editing and gene regulation in various organisms, but it can facilitate only one activity at a time within any given cell.
특징 및 장점
- Highly specific
- Highly active
- Ready-to-inject/transfect
포장
pkg of 50 μg (≥ 300 pmol)
pkg of 250 μg (≥ 1500 pmol)
pkg of 250 μg (≥ 1500 pmol)
성분
Each kit consists of:
- one vial of Cas9 recombinant protein
- one vial containing 1 mL of 1× dilution buffer
- one vial containing 1 mL of nuclease-free water with glycerol
원리
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 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.
재구성
Lyophilized S. pyogenes Cas9 protein should be resuspended in the Reconstitution solution provided to desired concentration. Gently tap tube to completely dissolve lyophilized powder, incubate for 10 minutes on ice, and spin tube to bring material to bottom of tube.
기타 정보
Use our CRISPR Selection Tool to order gRNA
Check out our other MISSION® Cas9 Proteins at SigmaAldrich.com/CRISPRproteins
Check out our other MISSION® Cas9 Proteins at SigmaAldrich.com/CRISPRproteins
법적 정보
MISSION is a registered trademark of Merck KGaA, Darmstadt, Germany
키트 구성품 전용
제품 번호
설명
- Cas9-NLS from Streptococcus pyogenes, expressed in Escherichia coli
- Dilution buffer for Cas9 proteins
- Reconstitution solution for Cas9 proteins
Storage Class Code
10 - Combustible liquids
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.
이미 열람한 고객
Yu Shirai et al.
Cell reports methods, 2(5), 100215-100215 (2022-06-01)
Current approaches for insect gene editing require microinjection of materials into early embryos. This severely limits the application of gene editing to a great number of insect species, especially to those whose reproduction systems preclude access to early embryos for
Joseph Andrew Whitley et al.
Journal of extracellular vesicles, 11(4), e12196-e12196 (2022-04-07)
CRISPR/Cas9 genome editing is a very promising avenue for the treatment of a variety of genetic diseases. However, it is still very challenging to encapsulate CRISPR/Cas9 machinery for delivery. Protein N-myristoylation is an irreversible co/post-translational modification that results in the
The Streptococcus thermophilus CRISPR/Cas system provides immunity in Escherichia coli
Sapranauskas R, et al.
Nucleic Acids Research, 39(21), 9275-9282 (2011)
Orthogonal Cas9 proteins for RNA-guided gene regulation and editing
Esvelt K M, et al.
Nature Methods, 10(11), 1116-1116 (2013)
Ana Carolina Seni-Silva et al.
BMC genomics, 23(1), 123-123 (2022-02-14)
The Natterin protein family was first discovered in the venom of the medically significant fish Thalassophryne nattereri, and over the last decade natterin-like genes have been identified in various organisms, notably performing immune-related functions. Previous findings support natterin-like genes as
프로토콜
Sigma-Aldrich® sgRNA, crRNA and tracrRNA come HPLC purified as standard with our performance guarantee—even for custom designs.
The CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) system was discovered in bacteria, where it functions as an adaptive immune system against invading viral and plasmid DNA.
Combine guaranteed sgRNAs with our comprehensive range of CRISPR products and tools, including Cas9 and delivery reagents, for efficient genome modification with higher specificity.
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