Evidence map›Paper›PMID 40263274›Full record

ArticleNature communications2025

Make-or-break prime editing for genome engineering in Streptococcus pneumoniae.

Monica Rengifo-Gonzalez, Maria-Vittoria Mazzuoli, Axel B Janssen, Anne-Stéphanie Rueff, Jessica Burnier, Xue Liu, Jan-Willem Veening

Abstract read
In one paragraph

Article in Nature communications, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.

0numbers the graph read from it
0cells of the map it votes in
8citing 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

8 citing papers in PubMed.

  1. Review
  2. Review
  3. Review
  4. Article
  5. Nanoluciferase-Based Reporters for Clostridioides difficile.Methods in molecular biology (Clifton, N.J.) · 2026
    Article
  6. Review
  7. Review
  8. Precision engineering of the probioticApplied and environmental microbiology · 2025
    Article
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

7 authors.

Monica Rengifo-Gonzalez *Department of Fundamental Microbiology, Faculty of Biology and Medicine, University of Lausanne, Biophore Building, CH-, Lausanne, Switzerland.
Maria-Vittoria Mazzuoli *Department of Fundamental Microbiology, Faculty of Biology and Medicine, University of Lausanne, Biophore Building, CH-, Lausanne, Switzerland.ORCID http://orcid.org/0000-0002-7071-1698
Axel B Janssen *Department of Fundamental Microbiology, Faculty of Biology and Medicine, University of Lausanne, Biophore Building, CH-, Lausanne, Switzerland.ORCID http://orcid.org/0000-0002-9865-447X
Anne-Stéphanie RueffDepartment of Fundamental Microbiology, Faculty of Biology and Medicine, University of Lausanne, Biophore Building, CH-, Lausanne, Switzerland.
Jessica BurnierDepartment of Fundamental Microbiology, Faculty of Biology and Medicine, University of Lausanne, Biophore Building, CH-, Lausanne, Switzerland.ORCID http://orcid.org/0009-0006-6187-8126
Xue LiuDepartment of Pathogen Biology, Base for International Science and Technology Cooperation: Carson Cancer Stem Cell Vaccines R&D Center, International Cancer Center, Shenzhen University Medical School, Shenzhen, Guangdong, China. xueliu@szu.edu.cn.ORCID http://orcid.org/0000-0001-6485-1865
Jan-Willem VeeningDepartment of Fundamental Microbiology, Faculty of Biology and Medicine, University of Lausanne, Biophore Building, CH-, Lausanne, Switzerland. Jan-Willem.Veening@unil.ch.ORCID http://orcid.org/0000-0002-3162-6634

Funding

EC | EU Framework Programme for Research and Innovation H2020 | H2020 Priority Excellent Science | H2020 European Research Council (H2020 Excellent Science - European Research Council) 771534-PneumoCaTChERSchweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (Swiss National Science Foundation) 310030_200792Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (Swiss National Science Foundation) NCCR AntiResist 51NF40_180541Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (Swiss National Science Foundation) TMPFP3_210202
6 · The paper itself

Abstract

CRISPR-Cas9 has revolutionized genome engineering by allowing precise introductions of DNA double-strand breaks (DSBs). However, genome engineering in bacteria is still a complex, multi-step process requiring a donor DNA template for repair of DSBs. Prime editing circumvents this need as the repair template is indirectly provided within the prime editing guide RNA (pegRNA). Here, we developed make-or-break Prime Editing (mbPE) that allows for precise and effective genetic engineering in the opportunistic human pathogen Streptococcus pneumoniae. In contrast to traditional prime editing in which a nicking Cas9 is employed, mbPE harnesses wild type Cas9 in combination with a pegRNA that destroys the seed region or protospacer adjacent motif. Since most bacteria poorly perform template-independent end joining, correctly genome-edited clones are selectively enriched during mbPE. We show that mbPE is RecA-independent and can be used to introduce point mutations, deletions and targeted insertions, including protein tags such as a split luciferase, at selection efficiencies of over 93%. mbPE enables sequential genome editing, is scalable, and can be used to generate pools of mutants in a high-throughput manner. The mbPE system and pegRNA design guidelines described here will ameliorate future bacterial genome editing endeavors.

Indexed as

CRISPR-Cas SystemsGene EditingGenetic EngineeringGenome, BacterialStreptococcus pneumoniaeCRISPR-Associated Protein 9DNA Breaks, Double-StrandedRNA, Guide, CRISPR-Cas SystemsCRISPR-Associated Protein 9RNA, Guide, CRISPR-Cas Systems

Identifiers

PMID40263274
PMCPMC12015366

What OpenQuestion holds

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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.