Evidence map›Paper›PMID 40764055›Full record

ArticleGenome research2025

3'-end ligation sequencing is a sensitive method to detect DNA nicks at potential sites of off-target activity induced by prime editors.

Jacob Stewart-Ornstein, Matthew J Irby, Marina K Lilieholm, Dylan Laprise, Maria D Collier, Thomas Aunins, Dewi Harjanto, Aaron N Chang, Deepak Reyon, Jeremy S Duffield

Abstract read
In one paragraph

Article in Genome research, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2 papers.

0numbers the graph read from it
0cells of the map it votes in
2citing 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

2 citing papers in PubMed.

  1. Prime Editing for p47The New England journal of medicine · 2026
    Trial
  2. A primer on prime: A prime editing update from advances to first-in-human trial.Molecular therapy : the journal of the American Society of Gene Therapy · 2026
    Review
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

10 authors.

Jacob Stewart-OrnsteinPrime Medicine Incorporated, Cambridge, Massachusetts 02141, USA jstewart-ornstein@primemedicine.com.ORCID 0000-0001-8255-5904
Matthew J IrbyPrime Medicine Incorporated, Cambridge, Massachusetts 02141, USA.
Marina K LilieholmPrime Medicine Incorporated, Cambridge, Massachusetts 02141, USA.
Dylan LaprisePrime Medicine Incorporated, Cambridge, Massachusetts 02141, USA.
Maria D CollierPrime Medicine Incorporated, Cambridge, Massachusetts 02141, USA.
Thomas AuninsPrime Medicine Incorporated, Cambridge, Massachusetts 02141, USA.
Dewi HarjantoPrime Medicine Incorporated, Cambridge, Massachusetts 02141, USA.
Aaron N ChangPrime Medicine Incorporated, Cambridge, Massachusetts 02141, USA.
Deepak ReyonPrime Medicine Incorporated, Cambridge, Massachusetts 02141, USA.
Jeremy S DuffieldPrime Medicine Incorporated, Cambridge, Massachusetts 02141, USA.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Gene editing makes precise changes in DNA to restore normal function or expression of genes; however, the advancement of gene editing to the clinic is limited by the potential genotoxicity of off-target editing. To comprehensively identify potential sites in the genome that may be recognized by gene editing agents, in vitro approaches, in which the editor is combined with human genomic DNA and sites where editing may occur are identified biochemically, are important tools. Existing biochemical approaches for off-target discovery recognize double-stranded breaks generated by nuclease-based gene editors such as SpCas9, but novel approaches are needed for new editing modalities, such as prime editing, that nick one strand of DNA. To fill this gap, we have developed 3'-end ligation sequencing (PEG-seq), which can identify prime editor-induced nicks throughout the genome on in vitro digested human genomic DNA to identify potential off-target sites. Here we show that PEG-seq is an important addition to the off-target detection toolkit, enabling off-target discovery for DNA nicking gene editors such as prime editors.

Indexed as

DNAGene EditingSequence Analysis, DNACRISPR-Cas SystemsDNA Breaks, Double-StrandedGenome, HumanHumansDNA

Identifiers

PMID40764055
PMCPMC12401060

What Socratic holds

Textmetadata
LicenceCC BY-NC
Read underepoch 390

Registered trials

None linked

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 Socratic graph.