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Flow Cytometric Assays for Interrogating LAGLIDADG Homing Endonuclease DNA-Binding and Cleavage Properties

 Last Update: 2020-12-16
 Source: Internet
 Author: User

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flow cytometry protocol

blood flow cytometry

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A fast, easy, and scalable method to assess the properties of site-specific nucleases is crucial to �understanding their
in cellulo
behavior in genome engineering or population-level gene drive applications. Here we describe an analytical platform thatenables high-throughput, semiquantitative interrogation of the
DNA
-binding and catalytic properties of LAGLIDADG homing endonucleases(LHEs). Using this platform, natural or engineered LHEs are expressed on the surface of
Saccharomyces cerevisiae
yeast where they can be rapidly evaluated against synthetic DNA target sequences using flow cytometry.

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