Evidence map›Paper›PMID 41414668›Full record

ArticleNucleic acids research2025

Characterization and engineering of a type IV-A3 CRISPR-Cas system for genome editing in Escherichia coli.

Szabolcs Semsey, Emilie Søndberg, Mathilde Røen, Björn Hallström, Anders Østergaard Petersen, Lauge Alfastsen, Berta Rosich Bosch, Benjamin Wohl, Jasper Clube, Eric van der Helm and 2 more

Abstract read
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Article in Nucleic acids research, 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

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

12 authors.

Szabolcs SemseySNIPR Biome ApS, 2100, Copenhagen, Denmark.
Emilie SøndbergSNIPR Biome ApS, 2100, Copenhagen, Denmark.
Mathilde RøenSNIPR Biome ApS, 2100, Copenhagen, Denmark.
Björn HallströmSNIPR Biome ApS, 2100, Copenhagen, Denmark.
Anders Østergaard PetersenSNIPR Biome ApS, 2100, Copenhagen, Denmark.
Lauge AlfastsenSNIPR Biome ApS, 2100, Copenhagen, Denmark.
Berta Rosich BoschSNIPR Biome ApS, 2100, Copenhagen, Denmark.
Benjamin WohlSNIPR Biome ApS, 2100, Copenhagen, Denmark.
Jasper ClubeSNIPR Biome ApS, 2100, Copenhagen, Denmark.
Eric van der HelmSNIPR Biome ApS, 2100, Copenhagen, Denmark.
Christian GroendahlSNIPR Biome ApS, 2100, Copenhagen, Denmark.
Ioannis MougiakosSNIPR Biome ApS, 2100, Copenhagen, Denmark.ORCID 0000-0001-8853-8208

Funding

SNIPR Biome
6 · The paper itself

Abstract

CRISPR-Cas systems have revolutionized genome engineering technologies, but type IV CRISPR-Cas systems and their genome engineering potential have been critically underexplored. In this study, we identified a type IV-A3 CRISPR-Cas system from a clinical Klebsiella pneumoniae isolate and characterized its plasmid targeting activity and capacity to suppress chromosomal and plasmid gene expression in Escherichia coli. We revealed the pivotal role of Csf3 (Cas5) and the dispensable roles of Csf1 (Cas8-like) and Csf4 (DinG helicase) subunits in IV-A3 CRISPR-Cas complex formation. The system prevents plasmid propagation via interplay between DinG helicase activity and strategic protospacer positioning relative to plasmid replication and maintenance components. We enabled the IV-A3 CRISPR-Cas system to introduce lethal, sequence-specific double-stranded (ds)DNA breaks in the E. coli chromosome by fusing the nuclease domain of the I-TevI nuclease to the Cas8 N-terminus. Further, we developed a series of base editors, with various editing efficiencies and windows, by fusing the PmCDA1 cytidine deaminase to the Cas8, Cas5, and DinG subunits. Finally, conjugative transfer of the Cas5-PmCDA1 base editor into E. coli deactivated the tryptophan repressor gene, boosting IAA production. Our study provides new insights into type IV-A3 CRISPR-Cas systems and highlights their potential in genome engineering applications.

Indexed as

CRISPR-Cas SystemsEscherichia coliGene EditingBacterial ProteinsDNA HelicasesGenome, BacterialKlebsiella pneumoniaePlasmidsBacterial ProteinsDNA Helicases

Identifiers

PMID41414668
PMCPMC12714566

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.