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

Andrew D. Johnston, Claudia A. Simões-Pires, Masako Suzuki, John M. Greally

Research output: Contribution to journalArticle

Abstract

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 <5–10% are frequently typical for many other cell types. The protocol does not require the use of lentiviruses or stable transfection, permitting lymphoblastoid cell lines to be used for CRISPR-mediated genomic targeting and screening in population genetic studies.

Original languageEnglish (US)
Article number312
JournalCommunications Biology
Volume2
Issue number1
DOIs
StatePublished - Dec 1 2019

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Clustered Regularly Interspaced Short Palindromic Repeats
Human herpesvirus 4
oligonucleotides
Human Herpesvirus 4
Viruses
Oligonucleotides
B-lymphocytes
B-Lymphocytes
Cells
Population Genetics
transfection
genomics
Transfection
population genetics
Cell Line
Lentivirus
Screening
human cell lines
cells
Genetic Recombination

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • Medicine (miscellaneous)

Cite this

High-efficiency genomic editing in Epstein-Barr virus-transformed lymphoblastoid B cells using a single-stranded donor oligonucleotide strategy. / Johnston, Andrew D.; Simões-Pires, Claudia A.; Suzuki, Masako; Greally, John M.

In: Communications Biology, Vol. 2, No. 1, 312, 01.12.2019.

Research output: Contribution to journalArticle

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