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Flow Induced Dispersion Analysis to measure gRNA-Nuclease Interactions for CRISPR-CAS Development
Kritika Ray Dr. rer. nat.; Linh B. Truong, Carl Co
This Fidabio poster, produced in collaboration with Arbor Biotechnologies, demonstrates how Flow Induced Dispersion Analysis (FIDA) was used to measure the hydrodynamic radius of guide RNA (gRNA) alone and bound to Cas nuclease, quantifying gRNA–Cas binding and ranking CRISPR-Cas guide-RNA candidates by affinity. Titrating Cas nuclease against labelled gRNA gave a sigmoidal binding curve (size rising from 2.3 to 4.5 nm), and a competition assay ranked unlabelled candidate gRNAs using only 40 nL of sample. It is an example of using FIDA to characterise protein–nucleic-acid binding and support CRISPR-Cas guide-RNA screening.
