Purdue Research Foundation

Kok Zhi Lee

Kevin V Solomon

Michael Harris

Yu-Hsuan Lee

Loretta Sue Loesch-Fries

This invention relates to a method to produce gene alterations in the genomes of a eukaryotic and prokaryotic host cell. The method comprises utilizing Argonaute from Natronobacterium gregoryi (NgAgo) or a mutant thereof, and complementary 5' phosphorylated single-stranded DNA that target the enzyme to cleave specific regions of the chromosome. Additionally, N-terminal truncations (deletion of the repA domain; N-del), or its mutants including N-del/E598A, N-del/D601P, and N-del/E602P reduces random cleavage and can be used for targeted gene editing with a guide DNA. An expression system or a host cell and method of creating thereof are also in the scope of this application.

NgAgo-based gene-editing method and the uses thereof

Methods and nucleic acid sequences for the synthesis of biotemplates in a non-plant based expression system are provided. Such biotemplates include Barley stripe mosaic virus viral-like particles (BSMV-VLPs) that are capable of self-assembly due to being operatively linked with an origin of self-assembly with the Barley stripe mosaic virus capsid protein (BSMV-CP). Also provided are BSMV-VLPs that are capable of self-assembly due one or more site-directed mutations on the BSMV-CP, and BSMV-VLPs that exhibit enhanced stability due to such site-directed mutation(s).

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West Lafayette, IN, United States of America

Kok Zhi Lee