Baculoviral delivery of CRISPR/Cas9 facilitates efficient genome editing in human cells

The CRISPR/Cas9 system is a highly effective tool for genome editing. Key to robust genome editing is the efficient delivery of the CRISPR/Cas9 machinery. Viral delivery systems are efficient vehicles for the transduction of foreign genes but commonly used viral vectors suffer from a limited capacit...

Full description

Saved in:
Bibliographic Details
Published in:PloS one 2017-06, Vol.12 (6), p.e0179514-e0179514
Main Authors: Hindriksen, Sanne, Bramer, Arne J, Truong, My Anh, Vromans, Martijn J M, Post, Jasmin B, Verlaan-Klink, Ingrid, Snippert, Hugo J, Lens, Susanne M A, Hadders, Michael A
Format: Article
Language:English
Subjects:
Amino acids
Baculoviridae - genetics
Baculovirus
Biology and Life Sciences
Biotechnology
Cancer
Cell Line, Tumor
Cell lines
Cells (Biology)
Cloning
Cohesin
Cohesion
CRISPR
CRISPR-Cas Systems - genetics
Deoxyribonucleic acid
DNA
Editing
Efficiency
Engineering and Technology
Fluorescence
Fragmentation
Fragments
Gene Editing - methods
Gene expression
Gene Knockout Techniques
Gene mutation
Genes
Genome editing
Genome, Human - genetics
Genomes
Genomics
Homology
Humans
Intracellular Signaling Peptides and Proteins - genetics
Kinases
Localization
Marking
Medical research
Medicine
Mutation
Phosphorylation
Point Mutation
Protein-Serine-Threonine Kinases - genetics
Proteomics
Repair
Research and Analysis Methods
Robustness
Tags
Transcription
Viruses
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The CRISPR/Cas9 system is a highly effective tool for genome editing. Key to robust genome editing is the efficient delivery of the CRISPR/Cas9 machinery. Viral delivery systems are efficient vehicles for the transduction of foreign genes but commonly used viral vectors suffer from a limited capacity in the genetic information they can carry. Baculovirus however is capable of carrying large exogenous DNA fragments. Here we investigate the use of baculoviral vectors as a delivery vehicle for CRISPR/Cas9 based genome-editing tools. We demonstrate transduction of a panel of cell lines with Cas9 and an sgRNA sequence, which results in efficient knockout of all four targeted subunits of the chromosomal passenger complex (CPC). We further show that introduction of a homology directed repair template into the same CRISPR/Cas9 baculovirus facilitates introduction of specific point mutations and endogenous gene tags. Tagging of the CPC recruitment factor Haspin with the fluorescent reporter YFP allowed us to study its native localization as well as recruitment to the cohesin subunit Pds5B.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0179514