Evidence map›Paper›PMID 41776280›Full record

ArticleCommunications biology2026

HUH-tagged Cas9 as a platform for efficient ssODN-mediated knock-in via embryo and adult injection in insects.

Yu Shirai, Jonchee A Kao, Tarun Kumar, Naoki Matsuda, Ryoya Nakagawa, Osamu Nureki, Cassandra G Extavour, Takaaki Daimon

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Communications biology, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

0numbers the graph read from it
0cells of the map it votes in
1citing 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

1 citing paper in PubMed.

  1. 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

8 authors.

Yu ShiraiDepartment of Applied Biosciences, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.ORCID http://orcid.org/0009-0009-0346-7873
Jonchee A KaoDepartment of Molecular and Cellular Biology, Harvard University, Cambridge, MA, USA.ORCID http://orcid.org/0000-0003-1701-3265
Tarun KumarDepartment of Organismic & Evolutionary Biology, Harvard University, Cambridge, MA, USA.ORCID http://orcid.org/0000-0003-4071-4342
Naoki MatsudaDepartment of Applied Biosciences, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.
Ryoya NakagawaDepartment of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan.
Osamu NurekiDepartment of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan.ORCID http://orcid.org/0000-0003-1813-7008
Cassandra G ExtavourDepartment of Organismic & Evolutionary Biology, Harvard University, Cambridge, MA, USA.ORCID http://orcid.org/0000-0003-2922-5855
Takaaki DaimonDepartment of Applied Biosciences, Graduate School of Agriculture, Kyoto University, Kyoto, Japan. daimon.takaaki.7a@kyoto-u.ac.jp.ORCID http://orcid.org/0000-0002-8382-1132

Funding

Cabinet Office, Government of Japan JPJ008000Human Frontier Science Program (HFSP) Long-Term FellowMEXT | Japan Society for the Promotion of Science (JSPS) 20H02999, 20K21311, 22K19179, 24H00511, 24K21869MEXT | Japan Society for the Promotion of Science (JSPS) 21J20658MEXT | Japan Society for the Promotion of Science (JSPS) Overseas Research FellowNSF | BIO | Division of Integrative Organismal Systems (IOS) IOS-2220747
6 · The paper itself

Abstract

Recent advances in adult injection-based insect genome editing have enabled genetic manipulation of a wide range of insect species, including those previously considered difficult or even impervious to genetic modification. However, achieving efficient knock-in remains a significant challenge with this approach. Here, we demonstrate that fusing a HUH endonuclease tag to Cas9 significantly enhances both non-homologous end joining (NHEJ)-mediated knockout and homology-directed repair (HDR)-mediated knock-in via adult injection. This fusion increased knockout efficiency by up to fivefold in the beetle Tribolium castaneum through adult injection, likely due to its previously unrecognized nuclear localization activity. It also improved single-stranded oligodeoxynucleotide (ssODN)-mediated knock-in efficiency, which we attribute to its characteristic ssDNA-tethering activity. To evaluate its versatility, we tested the HUH-tagged Cas9 in conventional embryo injection, which significantly enhanced HDR-mediated knock-in of an epitope tag in cricket and milkweed bug embryos. Our findings establish the HUH-tag as a versatile platform for improving both NHEJ- and HDR-based genome editing, providing a robust framework to advance genetic engineering across a broad spectrum of arthropods.

Indexed as

CRISPR-Associated Protein 9CRISPR-Cas SystemsGene EditingGene Knock-In TechniquesOligodeoxyribonucleotidesTriboliumAnimalsDNA End-Joining RepairDNA, Single-StrandedCRISPR-Associated Protein 9DNA, Single-StrandedOligodeoxyribonucleotides

Identifiers

PMID41776280
PMCPMC13065834

What OpenQuestion holds

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LicenceCC BY-NC-ND
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 OpenQuestion graph.