SpCas9 D10A Nickase (Active) is a Streptococcus pyogenes serotype M1 Full Length protein, in the 2 to 1368 aa range, expressed in Escherichia coli, with >=95% purity, < 10 EU/mg endotoxin level and suitable for SDS-PAGE, HPLC, Biological Activity.
Images
Key facts
- Purity
- >=95% SDS-PAGE
- Endotoxin level
- < 10 EU/mg
- Expression system
- Escherichia coli
- Tags
- His tag N-Terminus
- Applications
- SDS-PAGE, HPLC, Biological Activity
- Biologically active
- Yes
Amino acid sequence
D K K Y S I G L A I G T N S V G W A V I T D E Y K V P S K K F K V L G N T D R H S I K K N L I G A L L F D S G E T A E A T R L K R T A R R R Y T R R K N R I C Y L Q E I F S N E M A K V D D S F F H R L E E S F L V E E D K K H E R H P I F G N I V D E V A Y H E K Y P T I Y H L R K K L V D S T D K A D L R L I Y L A L A H M I K F R G H F L I E G D L N P D N S D V D K L F I Q L V Q T Y N Q L F E E N P I N A S G V D A K A I L S A R L S K S R R L E N L I A Q L P G E K K N G L F G N L I A L S L G L T P N F K S N F D L A E D A K L Q L S K D T Y D D D L D N L L A Q I G D Q Y A D L F L A A K N L S D A I L L S D I L R V N T E I T K A P L S A S M I K R Y D E H H Q D L T L L K A L V R Q Q L P E K Y K E I F F D Q S K N G Y A G Y I D G G A S Q E E F Y K F I K P I L E K M D G T E E L L V K L N R E D L L R K Q R T F D N G S I P H Q I H L G E L H A I L R R Q E D F Y P F L K D N R E K I E K I L T F R I P Y Y V G P L A R G N S R F A W M T R K S E E T I T P W N F E E V V D K G A S A Q S F I E R M T N F D K N L P N E K V L P K H S L L Y E Y F T V Y N E L T K V K Y V T E G M R K P A F L S G E Q K K A I V D L L F K T N R K V T V K Q L K E D Y F K K I E C F D S V E I S G V E D R F N A S L G T Y H D L L K I I K D K D F L D N E E N E D I L E D I V L T L T L F E D R E M I E E R L K T Y A H L F D D K V M K Q L K R R R Y T G W G R L S R K L I N G I R D K Q S G K T I L D F L K S D G F A N R N F M Q L I H D D S L T F K E D I Q K A Q V S G Q G D S L H E H I A N L A G S P A I K K G I L Q T V K V V D E L V K V M G R H K P E N I V I E M A R E N Q T T Q K G Q K N S R E R M K R I E E G I K E L G S Q I L K E H P V E N T Q L Q N E K L Y L Y Y L Q N G R D M Y V D Q E L D I N R L S D Y D V D H I V P Q S F L K D D S I D N K V L T R S D K N R G K S D N V P S E E V V K K M K N Y W R Q L L N A K L I T Q R K F D N L T K A E R G G L S E L D K A G F I K R Q L V E T R Q I T K H V A Q I L D S R M N T K Y D E N D K L I R E V K V I T L K S K L V S D F R K D F Q F Y K V R E I N N Y H H A H D A Y L N A V V G T A L I K K Y P K L E S E F V Y G D Y K V Y D V R K M I A K S E Q E I G K A T A K Y F F Y S N I M N F F K T E I T L A N G E I R K R P L I E T N G E T G E I V W D K G R D F A T V R K V L S M P Q V N I V K K T E V Q T G G F S K E S I L P K R N S D K L I A R K K D W D P K K Y G G F D S P T V A Y S V L V V A K V E K G K S K K L K S V K E L L G I T I M E R S S F E K N P I D F L E A K G Y K E V K K D L I I K L P K Y S L F E L E N G R K R M L A S A G E L Q K G N E L A L P S K Y V N F L Y L A S H Y E K L K G S P E D N E Q K Q L F V E Q H K H Y L D E I I E Q I S E F S K R V I L A D A N L D K V L S A Y N K H R D K P I R E Q A E N I I H L F T L T N L G A P A A F K Y F D T T I D R K R Y T S T K E V L D A T L I H Q S I T G L Y E T R I D L S Q L G G D
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application SDS-PAGE | Reactivity Reacts | Dilution info - | Notes - |
Application HPLC | Reactivity Reacts | Dilution info - | Notes - |
Application Biological Activity | Reactivity Reacts | Dilution info - | Notes - |
Target data
Function
CRISPR (clustered regularly interspaced short palindromic repeat) is an adaptive immune system that provides protection against mobile genetic elements (viruses, transposable elements and conjugative plasmids) (PubMed:21455174). CRISPR clusters contain spacers, sequences complementary to antecedent mobile elements, and target invading nucleic acids. CRISPR clusters are transcribed and processed into CRISPR RNA (crRNA). In type II CRISPR systems correct processing of pre-crRNA requires a trans-encoded small RNA (tracrRNA), endogenous ribonuclease 3 (rnc) and this protein. The tracrRNA serves as a guide for ribonuclease 3-aided processing of pre-crRNA; Cas9 only stabilizes the pre-crRNA:tracrRNA interaction and has no catalytic function in RNA processing (PubMed:24270795). Subsequently Cas9/crRNA/tracrRNA endonucleolytically cleaves linear or circular dsDNA target complementary to the spacer; Cas9 is inactive in the absence of the 2 guide RNAs (gRNA). The target strand not complementary to crRNA is first cut endonucleolytically, then trimmed 3'-5' exonucleolytically. DNA-binding requires protein and both gRNAs, as does nuclease activity. Cas9 recognizes the protospacer adjacent motif (PAM) in the CRISPR repeat sequences to help distinguish self versus nonself, as targets within the bacterial CRISPR locus do not have PAMs. DNA strand separation and heteroduplex formation starts at PAM sites; PAM recognition is required for catalytic activity (PubMed:24476820). Confers immunity against a plasmid with homology to the appropriate CRISPR spacer sequences (CRISPR interference) (PubMed:21455174).
Alternative names
csn1, SPy_1046, cas9, CRISPR-associated endonuclease Cas9/Csn1, SpCas9, SpyCas9
Recommended products
SpCas9 D10A Nickase (Active) is a Streptococcus pyogenes serotype M1 Full Length protein, in the 2 to 1368 aa range, expressed in Escherichia coli, with >=95% purity, < 10 EU/mg endotoxin level and suitable for SDS-PAGE, HPLC, Biological Activity.
Key facts
- Purity
- >=95% SDS-PAGE
- Endotoxin level
- < 10 EU/mg
- Expression system
- Escherichia coli
- Tags
- His tag N-Terminus
- Applications
- SDS-PAGE, HPLC, Biological Activity
- Biologically active
- Yes
- Biological activity
- in vitro cleavage activity data show that SpCas9 D10A Nickase can target and cleave more than 95% of the substrates.
- Accession
- Q99ZW2-1
- Animal free
- No
- Species
- Streptococcus pyogenes serotype M1
- Concentration
- Storage buffer
pH: 7.4
Constituents: 50% Glycerol (glycerin, glycerine), 1.752% Sodium chloride, 0.473% Tris HCl, 0.015% (R*,R*)-1,4-Dimercaptobutan-2,3-diol, 0.004% Ethylenediaminetetraacetic acid, disodium, dihydrate
Sequence info
Amino acid sequence
- D K K Y S I G L A I G T N S V G W A V I T D E Y K V P S K K F K V L G N T D R H S I K K N L I G A L L F D S G E T A E A T R L K R T A R R R Y T R R K N R I C Y L Q E I F S N E M A K V D D S F F H R L E E S F L V E E D K K H E R H P I F G N I V D E V A Y H E K Y P T I Y H L R K K L V D S T D K A D L R L I Y L A L A H M I K F R G H F L I E G D L N P D N S D V D K L F I Q L V Q T Y N Q L F E E N P I N A S G V D A K A I L S A R L S K S R R L E N L I A Q L P G E K K N G L F G N L I A L S L G L T P N F K S N F D L A E D A K L Q L S K D T Y D D D L D N L L A Q I G D Q Y A D L F L A A K N L S D A I L L S D I L R V N T E I T K A P L S A S M I K R Y D E H H Q D L T L L K A L V R Q Q L P E K Y K E I F F D Q S K N G Y A G Y I D G G A S Q E E F Y K F I K P I L E K M D G T E E L L V K L N R E D L L R K Q R T F D N G S I P H Q I H L G E L H A I L R R Q E D F Y P F L K D N R E K I E K I L T F R I P Y Y V G P L A R G N S R F A W M T R K S E E T I T P W N F E E V V D K G A S A Q S F I E R M T N F D K N L P N E K V L P K H S L L Y E Y F T V Y N E L T K V K Y V T E G M R K P A F L S G E Q K K A I V D L L F K T N R K V T V K Q L K E D Y F K K I E C F D S V E I S G V E D R F N A S L G T Y H D L L K I I K D K D F L D N E E N E D I L E D I V L T L T L F E D R E M I E E R L K T Y A H L F D D K V M K Q L K R R R Y T G W G R L S R K L I N G I R D K Q S G K T I L D F L K S D G F A N R N F M Q L I H D D S L T F K E D I Q K A Q V S G Q G D S L H E H I A N L A G S P A I K K G I L Q T V K V V D E L V K V M G R H K P E N I V I E M A R E N Q T T Q K G Q K N S R E R M K R I E E G I K E L G S Q I L K E H P V E N T Q L Q N E K L Y L Y Y L Q N G R D M Y V D Q E L D I N R L S D Y D V D H I V P Q S F L K D D S I D N K V L T R S D K N R G K S D N V P S E E V V K K M K N Y W R Q L L N A K L I T Q R K F D N L T K A E R G G L S E L D K A G F I K R Q L V E T R Q I T K H V A Q I L D S R M N T K Y D E N D K L I R E V K V I T L K S K L V S D F R K D F Q F Y K V R E I N N Y H H A H D A Y L N A V V G T A L I K K Y P K L E S E F V Y G D Y K V Y D V R K M I A K S E Q E I G K A T A K Y F F Y S N I M N F F K T E I T L A N G E I R K R P L I E T N G E T G E I V W D K G R D F A T V R K V L S M P Q V N I V K K T E V Q T G G F S K E S I L P K R N S D K L I A R K K D W D P K K Y G G F D S P T V A Y S V L V V A K V E K G K S K K L K S V K E L L G I T I M E R S S F E K N P I D F L E A K G Y K E V K K D L I I K L P K Y S L F E L E N G R K R M L A S A G E L Q K G N E L A L P S K Y V N F L Y L A S H Y E K L K G S P E D N E Q K Q L F V E Q H K H Y L D E I I E Q I S E F S K R V I L A D A N L D K V L S A Y N K H R D K P I R E Q A E N I I H L F T L T N L G A P A A F K Y F D T T I D R K R Y T S T K E V L D A T L I H Q S I T G L Y E T R I D L S Q L G G D
- Accession
- Q99ZW2
- Protein length
- Full Length
- Predicted molecular weight
- 163 kDa
- Amino acids
- 2 to 1368
- Nature
- Recombinant
- Tags
- His tag N-Terminus
Specifications
- Form
- Liquid
General info
- Function
CRISPR (clustered regularly interspaced short palindromic repeat) is an adaptive immune system that provides protection against mobile genetic elements (viruses, transposable elements and conjugative plasmids) (PubMed:21455174). CRISPR clusters contain spacers, sequences complementary to antecedent mobile elements, and target invading nucleic acids. CRISPR clusters are transcribed and processed into CRISPR RNA (crRNA). In type II CRISPR systems correct processing of pre-crRNA requires a trans-encoded small RNA (tracrRNA), endogenous ribonuclease 3 (rnc) and this protein. The tracrRNA serves as a guide for ribonuclease 3-aided processing of pre-crRNA; Cas9 only stabilizes the pre-crRNA:tracrRNA interaction and has no catalytic function in RNA processing (PubMed:24270795). Subsequently Cas9/crRNA/tracrRNA endonucleolytically cleaves linear or circular dsDNA target complementary to the spacer; Cas9 is inactive in the absence of the 2 guide RNAs (gRNA). The target strand not complementary to crRNA is first cut endonucleolytically, then trimmed 3'-5' exonucleolytically. DNA-binding requires protein and both gRNAs, as does nuclease activity. Cas9 recognizes the protospacer adjacent motif (PAM) in the CRISPR repeat sequences to help distinguish self versus nonself, as targets within the bacterial CRISPR locus do not have PAMs. DNA strand separation and heteroduplex formation starts at PAM sites; PAM recognition is required for catalytic activity (PubMed:24476820). Confers immunity against a plasmid with homology to the appropriate CRISPR spacer sequences (CRISPR interference) (PubMed:21455174).
- Sequence similarities
Belongs to the CRISPR-associated protein Cas9 family. Subtype II-A subfamily.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
- Storage information
- Avoid freeze / thaw cycle
This product is an active protein and may elicit a biological response in vivo, handle with caution.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
CRISPR-Cas9 also known simply as Cas9 is a protein that acts as a molecular scissor in gene editing. It has a molecular weight of approximately 160 kDa. Cas9 is a part of the CRISPR-Cas system originally discovered in bacteria where it serves as an adaptive immune system. In bacterial cells Cas9 targets and cleaves specific DNA sequences allowing for targeted gene modifications. While the expression of the CRISPR-Cas system occurs naturally in prokaryotes scientists now harness it in various organisms for genetic manipulation.
- Biological function summary
The Cas9 protein functions as an integral part of the CRISPR-Cas9 complex which includes a guide RNA to direct the protein to specific DNA sequences. This complex enables precise cuts at targeted locations within the genome. The Cas9 protein size allows it to fit effectively within cells facilitating genome editing in areas such as research agriculture and therapeutics. The complexity of CRISPR-Cas9 also includes the interaction with other cellular components that assist in DNA repair post-cleavage.
- Pathways
CRISPR-Cas9 plays a role in DNA repair pathways particularly non-homologous end joining and homologous recombination. After Cas9-induced DNA breaks these pathways become active to repair the cleaved DNA. The gene editing process facilitated by CRISPR-Cas9 often involves interaction with DNA repair proteins like ku70/80 and Rad51 in response to induced breaks. This allows for either the incorporation of new genetic material or the modification of existing genes.
- Associated diseases and disorders
CRISPR-Cas9 has potential in treating genetic disorders such as cystic fibrosis and Duchenne muscular dystrophy. By correcting mutated genes scientists aim to restore normal gene function offering potential therapeutic benefits. Additionally CRISPR-Cas9 relates indirectly to p53 a protein known for its tumor suppressor functions because gene-editing processes can sometimes activate p53-dependent pathways leading to cell cycle arrest or apoptosis. This highlights the importance of understanding the broader implications of CRISPR-Cas9 in therapeutic applications.
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3 product images
Biological Activity - SpCas9 D10A Nickase (Active) (ab316172)
90% of the supercoiled plasmid DNA can be cleaved into open circular plasmid DNA by 40-80ng Recombinant SpCas9 D10A Nickase in the 30 μl reaction system.
HPLC - SpCas9 D10A Nickase (Active) (ab316172)
The purity of SpCas9 D10A Nickase detected by SEC-HPLC is greater than 95%.
SDS-PAGE - SpCas9 D10A Nickase (Active) (ab316172)
The purity of Recombinant SpCas9 D10A Nickase detected by Bis-Tris PAGE is greater than 95%.
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