Here, the scientists use single-molecule imaging to visualize Cascade and Cas3 binding to foreign DNA targets. Their analysis reveals two distinct pathways dictated by the presence or absence of a protospacer-adjacent motif (PAM). Binding to a protospacer flanked by a PAM recruits a nuclease-active Cas3 for degradation of short single-stranded regions of target DNA, whereas PAM mutations elicit an alternative pathway that recruits a nuclease-inactive Cas3 through a mechanism that is dependent on the Cas1 and Cas2 proteins.
The authors introduce the Protospacer Workbench. This is an powerfull offline software for rapid, flexible design of Cas9 sgRNA with a smart graphical user interface
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Here, the scientists use single-molecule imaging to visualize Cascade and Cas3 binding to foreign DNA targets. Their analysis reveals two distinct pathways dictated by the presence or absence of a protospacer-adjacent motif (PAM). Binding to a protospacer flanked by a PAM recruits a nuclease-active Cas3 for degradation of short single-stranded regions of target DNA, whereas PAM mutations elicit an alternative pathway that recruits a nuclease-inactive Cas3 through a mechanism that is dependent on the Cas1 and Cas2 proteins.
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