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Bacterial CRISPR-Cas systems have recently been repurposed as RNA-guided genome editors in research labs across the world. Yet CRISPR-Cas adaptive immune systems are very diverse and many systems remain uncharacterized. Discovering the structure and function of newly discovered and uncharacterized systems may further advance existing genome editing technologies, or lead to new ones. To better understand the function of the little-researched Type IV-B system we cloned the genes of a Type IV-B system from Mycobacterium J623 into a plasmid containing a target sequence of a Type IV-A and Type V system. The Type IV-B system was placed on the target plasmid in order to test the hypothesis that Type IV-B acts as an anti-CRISPR system by binding up CRISPR derived RNAs before they can be bound by the Cas proteins of the immune system. Our cloning was found to be successful through DNA sequencing, providing a path towards testing our hypothesis, as well as future studies aimed at characterizing the structure and function of IV-B systems.

Publisher

Utah State University

Publication Date

12-10-2020

Disciplines

Chemistry | Physical Sciences and Mathematics

Cloning Type IV-B CRISPR System into a Plasmid

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Chemistry Commons

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