High-efficiency genomic editing in Epstein-Barr virus-transformed lymphoblastoid B cells using a single-stranded donor oligonucleotide strategy

While human lymphoblastoid cell lines represent a valuable resource for population genetic studies, they have usually been regarded as difficult for CRISPR-mediated genomic editing because of very inefficient DNA transfection and retroviral or lentiviral transduction in these cells, which becomes a...

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Bibliographic Details
Published in:Communications biology 2019-08, Vol.2 (1), p.312, Article 312
Main Authors: Johnston, Andrew D., Simões-Pires, Claudia A., Suzuki, Masako, Greally, John M.
Format: Article
Language:English
Subjects:
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B-Lymphocytes - metabolism
B-Lymphocytes - virology
Biology
Biomedical and Life Sciences
Cell Line, Transformed
Child
Clone Cells
CRISPR
CRISPR-Cas Systems - genetics
Editing
Epstein-Barr virus
Gene Editing
Gene Rearrangement - genetics
Genetic Loci
Genomes
Herpesvirus 4, Human - metabolism
Homology
Humans
Life Sciences
Lymphoblastoid cell lines
Lymphocytes B
Oligonucleotides
Oligonucleotides - metabolism
Population genetics
Population studies
Recombination
Transfection
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Summary:While human lymphoblastoid cell lines represent a valuable resource for population genetic studies, they have usually been regarded as difficult for CRISPR-mediated genomic editing because of very inefficient DNA transfection and retroviral or lentiviral transduction in these cells, which becomes a substantial problem when multiple constructs need to be co-expressed. Here we describe a protocol using a single-stranded donor oligonucleotide strategy for ‘scarless’ editing in lymphoblastoid cells, yielding 12/60 (20%) of clones with homology-directed recombination, when rates of
ISSN:2399-3642
2399-3642
DOI:10.1038/s42003-019-0559-3