Adapting Next-Generation Sequencing to <i>in Process</i> CRISPR-Cas9 Genome Editing of Recombinant <i>Ac</i>MNPV Vectors: From Shotgun to Tiled-Amplicon Sequencing.

ABSTRACT: The alphabaculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is the most commonly used virus in the Baculovirus Expression Vector System (BEVS) and has been utilized for the production of many human and veterinary biologics. AcMNPV has a large dsDNA genome that remains understudied, and relatively unmodified from the wild-type, especially considering how extensively utilized it is as an expression vector. Previously, our group utilized CRISPR-Cas9 genome engineering that revealed phenotypic changes when baculovirus genes are targeted using either co-expressed sgRNA or transfected sgRNA into a stable insect cell line that produced the Cas9 protein. Here, we describe a pipeline to sequence the recombinant AcMNPV expression vectors using shotg

SUBMITTER: Chakraborty M 

PROVIDER: S-EPMC11946314 | biostudies-literature | 2025 Mar

REPOSITORIES: biostudies-literature