Difference between revisions of "Team:NTU-Singapore/Proof"
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| − | <h1> | + | <h1>PI Domain description </h1> |
</div> | </div> | ||
<div class="well"> | <div class="well"> | ||
| − | <p> | + | <p>PI domain is the second largest domain in SpCas9 sequence and plays an essential role in binding to the target DNA sequence at its upstream PAM sequence and also scaffolding gRNA inside its complex structure. As a result, we realized it may not be easily achievable by deleting the whole PI region and yet keeping the normal functionality of dCas9. However, we also believed PI domain is a potential domain to be truncated because that is where the main difference is found between SpCas9 (370aa) and the much more compact SaCas9 (144aa) in terms of their sizes. Thus, we have made a homology comparison between SpCas9 and SaCas9, focusing on their structural differences in PI domain. Subsequently, the main homological difference in their sequences was identified from aa1230S – aa1316N which is only present in SpCas9, containing 5 alpha helices. This identified sequence was named as PI1 for the labeling of the subsequent experiments. |
| − | <p> | + | </p> |
| + | <p>Meanwhile, we have also looked at the evolutionary homology pattern of the variants species of Cas9 and found out there are another region which is less conserved as compared to the rest of PI domain. We then labeled it as PI2 for potential target to be truncated. The sequence of PI2 is from aa1140A to aa1191L, which consists of two beta sheets and two alpha helices. Hence, we have identified two target regions PI1 and PI2 for the truncation test. | ||
| + | </p> | ||
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Revision as of 08:09, 16 October 2016
Playing with Cas9 and Co.
sub-text
PI Domain description
PI domain is the second largest domain in SpCas9 sequence and plays an essential role in binding to the target DNA sequence at its upstream PAM sequence and also scaffolding gRNA inside its complex structure. As a result, we realized it may not be easily achievable by deleting the whole PI region and yet keeping the normal functionality of dCas9. However, we also believed PI domain is a potential domain to be truncated because that is where the main difference is found between SpCas9 (370aa) and the much more compact SaCas9 (144aa) in terms of their sizes. Thus, we have made a homology comparison between SpCas9 and SaCas9, focusing on their structural differences in PI domain. Subsequently, the main homological difference in their sequences was identified from aa1230S – aa1316N which is only present in SpCas9, containing 5 alpha helices. This identified sequence was named as PI1 for the labeling of the subsequent experiments.
Meanwhile, we have also looked at the evolutionary homology pattern of the variants species of Cas9 and found out there are another region which is less conserved as compared to the rest of PI domain. We then labeled it as PI2 for potential target to be truncated. The sequence of PI2 is from aa1140A to aa1191L, which consists of two beta sheets and two alpha helices. Hence, we have identified two target regions PI1 and PI2 for the truncation test.
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Maecenas sed diam eget risus varius blandit sit amet non magna. Lorem ipsum dolor sit amet, consectetur adipiscing elit. Praesent commodo cursus magna, vel scelerisque nisl consectetur et. Cras mattis consectetur purus sit amet fermentum. Duis mollis, est non commodo luctus, nisi erat porttitor ligula, eget lacinia odio sem nec elit. Aenean lacinia bibendum nulla sed consectetur.
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Maecenas sed diam eget risus varius blandit sit amet non magna. Lorem ipsum dolor sit amet, consectetur adipiscing elit. Praesent commodo cursus magna, vel scelerisque nisl consectetur et. Cras mattis consectetur purus sit amet fermentum. Duis mollis, est non commodo luctus, nisi erat porttitor ligula, eget lacinia odio sem nec elit. Aenean lacinia bibendum nulla sed consectetur.
header
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Maecenas sed diam eget risus varius blandit sit amet non magna. Lorem ipsum dolor sit amet, consectetur adipiscing elit. Praesent commodo cursus magna, vel scelerisque nisl consectetur et. Cras mattis consectetur purus sit amet fermentum. Duis mollis, est non commodo luctus, nisi erat porttitor ligula, eget lacinia odio sem nec elit. Aenean lacinia bibendum nulla sed consectetur.
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Maecenas sed diam eget risus varius blandit sit amet non magna. Lorem ipsum dolor sit amet, consectetur adipiscing elit. Praesent commodo cursus magna, vel scelerisque nisl consectetur et. Cras mattis consectetur purus sit amet fermentum. Duis mollis, est non commodo luctus, nisi erat porttitor ligula, eget lacinia odio sem nec elit. Aenean lacinia bibendum nulla sed consectetur.
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Maecenas sed diam eget risus varius blandit sit amet non magna. Lorem ipsum dolor sit amet, consectetur adipiscing elit. Praesent commodo cursus magna, vel scelerisque nisl consectetur et. Cras mattis consectetur purus sit amet fermentum. Duis mollis, est non commodo luctus, nisi erat porttitor ligula, eget lacinia odio sem nec elit. Aenean lacinia bibendum nulla sed consectetur.
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Maecenas sed diam eget risus varius blandit sit amet non magna. Lorem ipsum dolor sit amet, consectetur adipiscing elit. Praesent commodo cursus magna, vel scelerisque nisl consectetur et. Cras mattis consectetur purus sit amet fermentum. Duis mollis, est non commodo luctus, nisi erat porttitor ligula, eget lacinia odio sem nec elit. Aenean lacinia bibendum nulla sed consectetur.
header
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Maecenas sed diam eget risus varius blandit sit amet non magna. Lorem ipsum dolor sit amet, consectetur adipiscing elit. Praesent commodo cursus magna, vel scelerisque nisl consectetur et. Cras mattis consectetur purus sit amet fermentum. Duis mollis, est non commodo luctus, nisi erat porttitor ligula, eget lacinia odio sem nec elit. Aenean lacinia bibendum nulla sed consectetur.
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Maecenas sed diam eget risus varius blandit sit amet non magna. Lorem ipsum dolor sit amet, consectetur adipiscing elit. Praesent commodo cursus magna, vel scelerisque nisl consectetur et. Cras mattis consectetur purus sit amet fermentum. Duis mollis, est non commodo luctus, nisi erat porttitor ligula, eget lacinia odio sem nec elit. Aenean lacinia bibendum nulla sed consectetur.
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Maecenas sed diam eget risus varius blandit sit amet non magna. Lorem ipsum dolor sit amet, consectetur adipiscing elit. Praesent commodo cursus magna, vel scelerisque nisl consectetur et. Cras mattis consectetur purus sit amet fermentum. Duis mollis, est non commodo luctus, nisi erat porttitor ligula, eget lacinia odio sem nec elit. Aenean lacinia bibendum nulla sed consectetur.
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Maecenas sed diam eget risus varius blandit sit amet non magna. Lorem ipsum dolor sit amet, consectetur adipiscing elit. Praesent commodo cursus magna, vel scelerisque nisl consectetur et. Cras mattis consectetur purus sit amet fermentum. Duis mollis, est non commodo luctus, nisi erat porttitor ligula, eget lacinia odio sem nec elit. Aenean lacinia bibendum nulla sed consectetur.
