Sandbox Reserved 1750: Difference between revisions

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The SaCas9 recognizes the sgRNA scaffold within the <scene name='92/925538/Lobes_and_linkers/20'>REC lobe and WED domain</scene>. The WED contains five stranded beta sheets flanked with four alpha helices to allow binding of the repeat: anti-repeat duplex. REC lob binds the scaffold and secures it into the SaCas9 <ref name="Cas9" />.

== Endonuclease Activity of Cas9 ==

Finally, <scene name='92/925538/Lobes_and_linkers/12'>RuvC and HNH</scene> are in endonuclease activity. RuvC uses two manganese to cleave the non-target DNA through manganese coordinating with the phosphate backbone and aspartic acid residues. A histidine then acts as a base to create a hydroxide nucleophile that attacks the phosphate bond and cleaves the non-target DNA. The binding of the RuvC to the target DNA changes the conformation of a linker protein region between the RuvC domain and the HNH domain. The conformational change of the linker bings the HNH domain close enough to the target DNA to cut the DNA. The HNH follows a similar mechanism as to RuvC using a histidine base to create a hydroxide ion nucleophile that attacks the phosphate bond. This is modeled as manganese however, it is often magnesium in cell <ref name="Cas9" />.

Finally, <scene name='92/925538/Lobes_and_linkers/12'>RuvC and HNH</scene> are involved in endonuclease activity. RuvC uses two manganese to cleave the non-target DNA through manganese coordinating with the phosphate backbone and aspartic acid residues. A histidine then acts as a base to create a hydroxide nucleophile that attacks the phosphate bond and cleaves the non-target DNA. The binding of the RuvC to the target DNA changes the conformation of a linker protein region between the RuvC domain and the HNH domain. The conformational change of the linker bings the HNH domain close enough to the target DNA to cut the DNA. The HNH follows a similar mechanism as to RuvC using a histidine base to create a hydroxide ion nucleophile that attacks the phosphate bond. This is modeled as manganese however, it is often magnesium in cell <ref name="Cas9" />.

</structuresection>

==References==

Sandbox Reserved 1750: Difference between revisions

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@@ Line 12: / Line 12: @@
 The SaCas9 recognizes the sgRNA scaffold within the <scene name='92/925538/Lobes_and_linkers/20'>REC lobe and WED domain</scene>. The WED contains five stranded beta sheets flanked with four alpha helices to allow binding of the repeat: anti-repeat duplex. REC lob binds the scaffold and secures it into the SaCas9 <ref name="Cas9" />.
 == Endonuclease Activity of Cas9 ==
-Finally, <scene name='92/925538/Lobes_and_linkers/12'>RuvC and HNH</scene> are in endonuclease activity. RuvC uses two manganese to cleave the non-target DNA through manganese coordinating with the phosphate backbone and aspartic acid residues. A histidine then acts as a base to create a hydroxide nucleophile that attacks the phosphate bond and cleaves the non-target DNA. The binding of the RuvC to the target DNA changes the conformation of a linker protein region between the RuvC domain and the HNH domain. The conformational change of the linker bings the HNH domain close enough to the target DNA to cut the DNA. The HNH follows a similar mechanism as to RuvC using a histidine base to create a hydroxide ion nucleophile that attacks the phosphate bond. This is modeled as manganese however, it is often magnesium in cell <ref name="Cas9" />.
+Finally, <scene name='92/925538/Lobes_and_linkers/12'>RuvC and HNH</scene> are involved in endonuclease activity. RuvC uses two manganese to cleave the non-target DNA through manganese coordinating with the phosphate backbone and aspartic acid residues. A histidine then acts as a base to create a hydroxide nucleophile that attacks the phosphate bond and cleaves the non-target DNA. The binding of the RuvC to the target DNA changes the conformation of a linker protein region between the RuvC domain and the HNH domain. The conformational change of the linker bings the HNH domain close enough to the target DNA to cut the DNA. The HNH follows a similar mechanism as to RuvC using a histidine base to create a hydroxide ion nucleophile that attacks the phosphate bond. This is modeled as manganese however, it is often magnesium in cell <ref name="Cas9" />.
 </structuresection>
 ==References==
 <references />