Cited by 1,675CRISPR and Genetic EngineeringRNA regulation and diseaseRNA Interference and Gene Delivery

Base editing: precision chemistry on the genome and transcriptome of living cells

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Rees, Holly A., Liu, David R.·2018·Nature Reviews Genetics

Abstract

RNA-guided programmable nucleases from CRISPR systems generate precise breaks in DNA or RNA at specified positions. In cells, this activity can lead to changes in DNA sequence or RNA transcript abundance. Base editing is a newer genome-editing approach that uses components from CRISPR systems together with other enzymes to directly install point mutations into cellular DNA or RNA without making double-stranded DNA breaks. DNA base editors comprise a catalytically disabled nuclease fused to a nucleobase deaminase enzyme and, in some cases, a DNA glycosylase inhibitor. RNA base editors achieve analogous changes using components that target RNA. Base editors directly convert one base or base pair into another, enabling the efficient installation of point mutations in non-dividing cells without generating excess undesired editing by-products. In this Review, we summarize base-editing strategies to generate specific and precise point mutations in genomic DNA and RNA, highlight recent developments that expand the scope, specificity, precision and in vivo delivery of base editors and discuss limitations and future directions of base editing for research and therapeutic applications.

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  • Base editors act only within a small window of the single-stranded R-loop that is generated upon Cas9 target recognition.

    “base editors act only within a small window of the single-stranded R-loop that is generated upon Cas9 target recognition30.”

    From One-step generation of tumor models by base editor multiplexing in adult stem cell-derived organoids (2023) · cites this paper as [30]

    The Introduction says Cas targeting creates a single-stranded R-loop whose bases can be modified by the editor. “Editing window and bystander edits” describes an approximately five-nucleotide window of efficient BE3 editing, but explicitly says bases outside that window can still be edited at lower efficiency. Thus, “only” overstates the restriction. (LLM, conf=0.88)

    Experimental — accuracy not yet measured
  • Base editing does not induce a high level of double-stranded DNA breaks.

    “A new-generation of genome editing tool, Base Editing (BE) system, enables precise base changes (reviewed in 4, 5), does not induce a high-level of DSBs, and finally does not require HDR donors for gene repair 4, 5.”
  • Base editing does not require homology-directed repair donors for gene repair.

    “A new-generation of genome editing tool, Base Editing (BE) system, enables precise base changes (reviewed in 4, 5), does not induce a high-level of DSBs, and finally does not require HDR donors for gene repair 4, 5.”
  • Unlike gene augmentation, editing tools are not limited by gene size, making them applicable to any gene, regardless of their inheritance pattern.

    “Unlike gene augmentation, editing tools are not limited by gene size, making them applicable to any gene, regardless of their inheritance pattern. 53 , 54”
  • ABEs are more commonly used because the G>A variant is the most prevalent in human diseases.

    “ABEs are more commonly used because the G > A variant is the most prevalent in human diseases. 54”

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Authors
Rees, H. A., & Liu, D. R.
Affiliations
Broad Institute (US)
Corresponding author
Liu, David R.
Venue
Nature Reviews Genetics
Published
2018-10-15
Volume
19
Issue
12
Pages
770-788
ISSN
  • 1471-0056 (ISSN-L)
  • 1471-0064
OpenAlex
W2897295193
Cited by
1,675
References
168
Citations by year
2026: 168 · 2025: 212 · 2024: 220 · 2023: 210 · 2022: 253 · 2021: 241 · 2020: 252 · 2019: 112 · 2018: 6
FWCI
15.23
Type
journal article
Language
en
Open access
Locations
  • Nature Reviews Genetics (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • Howard Hughes Medical Institute
  • National Human Genome Research Institute (RM1HG009490)
  • National Institute of General Medical Sciences (R01GM065865, R01GM065400, R35GM118062)
  • National Institute of Biomedical Imaging and Bioengineering (R01EB022376)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
Keywords
base editing, CRISPR, point mutations, in vivo delivery, genome editing, off-target editing, non-dividing cells, gene therapy
MeSH terms
Gene Editing, Animals, Endonucleases, Humans, DNA Breaks, Double-Stranded, Transcriptome, CRISPR-Cas Systems

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