Cited by 1,233CRISPR and Genetic EngineeringRNA regulation and diseaseCAR-T cell therapy research

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Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants

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Walton, Russell T., Christie, Kathleen A., Whittaker, Madelynn N., Kleinstiver, Benjamin P.·2020·Science

DOI 10.1126/science.aba8853PMC PMC7297043

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Abstract

A PAMless base editor: CRISPR-Cas DNA base editing typically requires a specific motif for targeting known as a protospacer-adjacent motif (PAM). This requirement limits the sequences within a genome that can be targeted. Walton et al. engineered specific variants of the Streptococcus pyogenes Cas9 enzyme named SpG and SpRY that could recognize and edit a wider array of PAMs. Using SpRY, the authors were able to correct previously uneditable mutations associated with human disease. Although off-target effects were observed for these engineered Cas enzymes at levels similar to those of the wild-type enzyme, depending on the context, these engineered enzymes widen the potential applications of precision genome editing. Science , this issue p. 290

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Authors: Walton, R. T., Christie, K. A., Whittaker, M. N., & Kleinstiver, B. P.
Affiliations: Massachusetts General Hospital (US)
Harvard University (US)
Corresponding author: Kleinstiver, Benjamin P.
ORCID: Walton, Russell T.
Christie, Kathleen A.
Whittaker, Madelynn N.
Kleinstiver, Benjamin P.
Venue: Science
Publisher: American Association for the Advancement of Science (AAAS)
Published: 2020-04-17
Volume: 368
Issue: 6488
Pages: 290-296
ISSN: 0036-8075 (ISSN-L)
1095-9203
DOI: 10.1126/science.aba8853
PMC: PMC7297043
OpenAlex: W3013504322
Semantic Scholar: 84609b61a82a11e3122a81b5bdae20a2822e577b
Cited by: 1,233
References: 48
Citations by year: 2026: 157 · 2025: 200 · 2024: 190 · 2023: 213 · 2022: 203 · 2021: 177 · 2020: 93
FWCI: 56.07
Type: journal article
Language: en
Open access: green
www.ncbi.nlm.nih.gov
Locations: Science (publishedVersion)
PubMed (publishedVersion)
PubMed Central (acceptedVersion)
Funders: National Heart, Lung, and Blood Institute (P01-HL142494)
National Cancer Institute (R00-CA218870)
American Society of Gene and Cell Therapy
Margaret Q. Landenberger Research Foundation
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
Health Sciences › Medicine › Oncology › CAR-T cell therapy research
Keywords: CRISPR/Cas9, SpCas9, SPG, SpRY, base editing, genome editing, human cells
MeSH terms: Protein Domains, Gene Editing, CRISPR-Associated Protein 9, Humans, Substrate Specificity, Mutagenesis, Gene Targeting, Genetic Predisposition to Disease, HEK293 Cells, CRISPR-Cas Systems

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Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants

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