Team:Waterloo/Modeling/PAM Flexibility

Modelling Engineered Cas9 PAM Flexibility

Copy and past code from this page as a template for the rest of the site

Why is it important to do this? / Is it possible

PAM Sites Differ between Natural Cas9 PRoteins

Engineered SpyCas9 NGAG and NGA

Computational Engineering of PAM Flexibility

Modelling Binding with PyMOL and PyRosetta

Canonical molecular dynamics data, mention source of PDB

Mutating PAM sites within PDB

Available PAM Affinity Data

Mention Klienstiver and

Engineering Pipeline

Summarize whole approach

Model Validation

Choice of DNA Mutation Software

Chimera VS 3DNA, give stats details

Patterns of Variation between Simulations

Simulations are Monte Carlo, we analyzed the differences between them

Wild-Type Cas9 PAM Affinities are Reproduced

Report Kendall's Tau, Pearson, Clustering Results

Mutant Cas9 PAM Affinities show Mixed Results

Mutating VQR/EQR, how EQR and VQR mutants change Cas9 visualizations, measures of distance from DNA bases to binding sites in the Cas9

Framework and Future Work

Proposed Tool

Link to software page re:pyrosetta

Future Work

Talk about adding other info like Gibb's Free Energy

Show a few graphs from Kleinstiver and link to dataset encouraging others to use

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