cas protein การใช้

• Class 2 systems use a single large Cas protein for the same purpose.
• CrRNAs associate with Cas proteins to form ribonucleotide complexes that recognize foreign nucleic acids.
• In vertebrates, the CAS protein family contains four members : EFS and CASS4.
• Class 1 systems use a complex of multiple Cas proteins to degrade foreign nucleic acids.
• Type III systems, like type I require six or seven Cas proteins binding to crRNAs.
• Evolutionary divergence of the CAS proteins family members is discussed by Singh et al . in detail.
• This  priming requires the Cas proteins involved in both acquisition and interference to interact with each other.
• Class 1 uses several Cas proteins together with the CRISPR RNAs ( crRNA ) to build a functional endonuclease.
• The type I-E complex ( commonly referred to as Cascade ) requires five Cas proteins bound to a single crRNA.
• These Cas proteins have a high structural homology, characterized by the presence of multiple protein interaction domains and phosphorylation motifs through which Cas family members can recruit effector proteins.
• Stage 3 : CRISPR interference . crRNA with a spacer that has strong complementarity to the incoming foreign DNA begins a cleavage event ( depicted with scissors ), which requires Cas proteins.
• When this CRISPR region of DNA is expressed by the cell, the small RNAs produced from the exogenous DNA inserts serve as a template sequence that other Cas proteins use to silence this same exogenous sequence.
• Class 2 CRISPR systems use a single Cas protein with a crRNA . Cpf1 has been recently identified as a Class II, Type V CRISPR / Cas systems containing a 1, 300 amino acid protein.
• RNA harboring the spacer sequence helps Cas proteins recognize and cut exogenous DNA . Other RNA-guided Cas proteins cut foreign RNA . CRISPRs are found in approximately 40 % of sequenced bacterial genomes and 90 % of sequenced archaea.
• RNA harboring the spacer sequence helps Cas proteins recognize and cut exogenous DNA . Other RNA-guided Cas proteins cut foreign RNA . CRISPRs are found in approximately 40 % of sequenced bacterial genomes and 90 % of sequenced archaea.
• Pre-crRNA is transcribed starting at the leader region by the host RNA polymerase and then cleaved by Cas proteins into smaller crRNAs containing a single spacer and a partial repeat ( shown as hairpin structure with colored spacers ).
• In 2008, Brouns and colleagues identified a complex of Cas protein that in " E . coli " cut the CRISPR RNA within the repeats into spacer-containing RNA molecules, which remained bound to the protein complex.
• The SH3 domain is a point of contact with polyproline sequences on focal adhesion kinase ( PTP-PEST proline-rich sequence 332 PPKPPR 337 has been shown to interact directly with the SH3 domain of members of EFS and another CAS protein, NEDD9.
• Type II systems use a trans-activating ( tracr ) RNA to form dsRNA, which is cleaved by Cas9 and RNaseIII . Type III systems use a Cas6 homolog that does not require hairpin loops in the direct repeat for cleavage . ( 6 ) In type II and type III systems secondary trimming is performed at either the 5 or 3 end to produce mature crRNAs . ( 7 ) Mature crRNAs associate with Cas proteins to form interference complexes . ( 8 ) In type I and type II systems, interactions between the protein and PAM sequence are required for degradation of invading DNA . Type III systems do not require a PAM for successful degradation and in type III-A systems basepairing occurs between the crRNA and mRNA rather than the DNA, targeted by type III-B systems.