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Targeting specificity of APOBEC-based cytosine base editor in human iPSCs determined by whole genome sequencing
- Source :
- Nature Communications, Nature Communications, Vol 10, Iss 1, Pp 1-9 (2019)
- Publication Year :
- 2019
-
Abstract
- DNA base editors have enabled genome editing without generating DNA double strand breaks. The applications of this technology have been reported in a variety of animal and plant systems, however, their editing specificity in human stem cells has not been studied by unbiased genome-wide analysis. Here we investigate the fidelity of cytidine deaminase-mediated base editing in human induced pluripotent stem cells (iPSCs) by whole genome sequencing after sustained or transient base editor expression. While base-edited iPSC clones without significant off-target modifications are identified, this study also reveals the potential of APOBEC-based base editors in inducing unintended point mutations outside of likely in silico-predicted CRISPR-Cas9 off-targets. The majority of the off-target mutations are C:G->T:A transitions or C:G->G:C transversions enriched for the APOBEC mutagenesis signature. These results demonstrate that cytosine base editor-mediated editing may result in unintended genetic modifications with distinct patterns from that of the conventional CRISPR-Cas nucleases.<br />Cytidine base editors are powerful tools for making subtle genome alterations. Here the authors analyse edited human iPSCs with whole genome sequencing and reveal the spectrum of off-target effects.
- Subjects :
- 0301 basic medicine
APOBEC
CRISPR-Cas9 genome editing
Science
Induced Pluripotent Stem Cells
General Physics and Astronomy
Computational biology
Biology
medicine.disease_cause
Genome
General Biochemistry, Genetics and Molecular Biology
Article
03 medical and health sciences
chemistry.chemical_compound
Cytosine
0302 clinical medicine
Genome editing
Cytidine Deaminase
Plant Cells
medicine
Animals
Humans
DNA Breaks, Double-Stranded
APOBEC Deaminases
lcsh:Science
Whole genome sequencing
Gene Editing
Mutation
Multidisciplinary
Whole Genome Sequencing
Genome, Human
Point mutation
Reproducibility of Results
General Chemistry
Stem-cell research
030104 developmental biology
chemistry
lcsh:Q
CRISPR-Cas Systems
030217 neurology & neurosurgery
DNA
Subjects
Details
- ISSN :
- 20411723
- Volume :
- 10
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Nature communications
- Accession number :
- edsair.doi.dedup.....765264fa7c516a48769f6fbf0436d9e6