An improved CRISPR-Cas9 protein-based method for knocking out insect Sf9 cell genes.

ABSTRACT: Insect cells are one of the uprising expression systems in the biopharmaceutical industry to produce vaccines and gene therapy vectors, but cell line development has been limited by the lack of established genetic engineering tools and genomic characterization. CRISPR-Cas9 has arisen as a powerful tool for gene editing but has seen little application in insect cells. In this work, a gene editing pipeline for the delivery of a ribonucleoprotein (RNP) complex comprised of a guide RNA and the enzyme Cas9 to insect Sf9 cells was implemented and then applied to knockout caspase initiator Sf-Dronc, aiming at alleviating cell apoptosis during an infection process. The resulting engineered cell lines were characterized as per their phenotype and production of three different product modalities. Ut

SUBMITTER: Graca M 

PROVIDER: S-EPMC12847097 | biostudies-literature | 2026 Jan

REPOSITORIES: biostudies-literature