Evidence map›Paper›PMID 41573864›Full record

ArticlebioRxiv : the preprint server for biology2025

A guide RNA repeat checkpoint steers CRISPR-Cas9 catalysis.

Ramadevi Chilamkurthy, Sruthi Sudhakar, Adrian A Pater, Michael S Bosmeny, Francis Stabile, Adam Katolik, S Harikrishna, Rolf Turk, Masad J Damha, Sabrina Leslie and 3 more

Abstract readPreprint
PubMed Full text at PMC Publisher
In one paragraph

Article in bioRxiv : the preprint server for biology, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

0numbers the graph read from it
0cells of the map it votes in
0citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

0 citing papers in PubMed.

No citing paper in PubMed yet.

4 · The record

Corrections and comments

5 · Who and what money

Authors and funding

13 authors.

Ramadevi ChilamkurthyDepartment of Biochemistry, Wake Forest University School of Medicine, Winston-Salem, NC, USA.
Sruthi SudhakarDepartment of Chemistry, Indian Institute of Technology Bombay, Powai, Mumbai, India.
Adrian A PaterDepartment of Biochemistry, Wake Forest University School of Medicine, Winston-Salem, NC, USA.
Michael S BosmenyDepartment of Biochemistry, Wake Forest University School of Medicine, Winston-Salem, NC, USA.
Francis StabileDepartment of Physics, McGill University, Montreal, Quebec, Canada.
Adam KatolikDepartment of Chemistry, McGill University, Montreal, Quebec, Canada.
S HarikrishnaDepartment of Chemistry, Indian Institute of Technology Bombay, Powai, Mumbai, India.
Rolf TurkIntegrated DNA Technologies, Coralville, Iowa, USA.
Masad J DamhaDepartment of Chemistry, McGill University, Montreal, Quebec, Canada.
Sabrina LeslieDepartment of Physics, McGill University, Montreal, Quebec, Canada.
Sergey KorolevDepartment of Biochemistry and Molecular Biology, School of Medicine, Saint Louis University, St. Louis, Missouri, USA.ORCID 0000-0001-9313-7126
P I PradeepkumarDepartment of Chemistry, Indian Institute of Technology Bombay, Powai, Mumbai, India.
Keith T GagnonDepartment of Biochemistry, Wake Forest University School of Medicine, Winston-Salem, NC, USA.

Funding

Nucleic Acid-Based Anti-CRISPR Inhibitors of Cas9R01GM135646 · NIGMS · WAKE FOREST UNIVERSITY HEALTH SCIENCES · PI GAGNON, KEITH THOMAS · 2020 to 2023
$1.1M
Mechanism of homologous recombination driven by the intrinsically disordered domain of tumor suppressor PALB2R01GM154323 · NIGMS · SAINT LOUIS UNIVERSITY · PI SERGEY KOROLEV · 2024 to 2026
$1.1M
NIGMS NIH HHS R01 GM135646NIGMS NIH HHS R01 GM154323
6 · The paper itself

Abstract

A widely adopted CRISPR-Cas9 modification is fusion of the naturally occurring two-component dual guide RNA (dgRNA) to create an artificial single guide RNA (sgRNA). However, mechanistic and functional differences between dgRNA and sgRNA have not been systematically explored. By investigating the activity of these two guide architectures, we discover a guide RNA repeat checkpoint (GRC) that senses the structure and dynamics of the guide repeat region. The GRC coordinates with other checkpoint mechanisms known to recognize spacer-target R-loop fidelity, together licensing target cleavage. Based on these principles, dgRNA and sgRNA properties could be combined into guide repeat-truncated sgRNAs (grtRNAs) and paired with high-fidelity Cas9 variants to further reduce off target editing. A mechanism that helps govern Cas9 catalysis via guide RNA structure sensing and communicates with other checkpoints offers a previously unappreciated path for understanding and further improving gene editing outcomes.

Indexed as

Cas9conformationcryo-EMdgRNAdual guideeditingGRCgrtRNAhigh-fidelitymolecular dynamicsoff targetRNAsgRNAsingle guidestructure

Identifiers

PMID41573864
PMCPMC12822677

What OpenQuestion holds

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LicenceCC BY-NC-ND
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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.