Evidence map›Paper›PMID 41814651›Full record

ArticleMolecular therapy : the journal of the American Society of Gene Therapy2026

Structural mining and engineering of metagenome-derived Cas12a orthologs expands the CRISPR genome editing and multiplex diagnostics toolkit.

Dagang Tao, Bingrong Xu, Sheng Li, Hailong Liu, Yinghui Wei, Xi Cao, Suyu Shi, Yuan Wang, Rui Jiang, Yonghui Zhang and 7 more

Abstract read
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In one paragraph

Article in Molecular therapy : the journal of the American Society of Gene Therapy, 2026. 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

17 authors.

Dagang TaoKey Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education & Key Lab of Swine Genetics and Breeding, Ministry of Agriculture and Rural Affairs, Huazhong Agricultural University, Wuhan 430070, P.R. China; The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan 430070, P.R. China.
Bingrong XuKey Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education & Key Lab of Swine Genetics and Breeding, Ministry of Agriculture and Rural Affairs, Huazhong Agricultural University, Wuhan 430070, P.R. China; Yazhouwan National Laboratory, Sanya 572024, P.R. China.
Sheng LiKey Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education & Key Lab of Swine Genetics and Breeding, Ministry of Agriculture and Rural Affairs, Huazhong Agricultural University, Wuhan 430070, P.R. China; The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan 430070, P.R. China.
Hailong LiuKey Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education & Key Lab of Swine Genetics and Breeding, Ministry of Agriculture and Rural Affairs, Huazhong Agricultural University, Wuhan 430070, P.R. China; The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan 430070, P.R. China.
Yinghui WeiHainan Institute, Northwest A&F University, Sanya 572024, P.R. China.
Xi CaoHainan Institute, Northwest A&F University, Sanya 572024, P.R. China.
Suyu ShiKey Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education & Key Lab of Swine Genetics and Breeding, Ministry of Agriculture and Rural Affairs, Huazhong Agricultural University, Wuhan 430070, P.R. China; The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan 430070, P.R. China.
Yuan WangKey Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education & Key Lab of Swine Genetics and Breeding, Ministry of Agriculture and Rural Affairs, Huazhong Agricultural University, Wuhan 430070, P.R. China; Yazhouwan National Laboratory, Sanya 572024, P.R. China.
Rui JiangKey Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education & Key Lab of Swine Genetics and Breeding, Ministry of Agriculture and Rural Affairs, Huazhong Agricultural University, Wuhan 430070, P.R. China; The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan 430070, P.R. China.
Yonghui ZhangKey Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education & Key Lab of Swine Genetics and Breeding, Ministry of Agriculture and Rural Affairs, Huazhong Agricultural University, Wuhan 430070, P.R. China; The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan 430070, P.R. China.
Changzhi ZhaoYazhouwan National Laboratory, Sanya 572024, P.R. China.
Jinxue RuanKey Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education & Key Lab of Swine Genetics and Breeding, Ministry of Agriculture and Rural Affairs, Huazhong Agricultural University, Wuhan 430070, P.R. China; The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan 430070, P.R. China.
Liangliang FuKey Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education & Key Lab of Swine Genetics and Breeding, Ministry of Agriculture and Rural Affairs, Huazhong Agricultural University, Wuhan 430070, P.R. China; The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan 430070, P.R. China.
Xingxu HuangLaboratory of Pancreatic Disease, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310058, P.R. China.
Xinyun LiKey Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education & Key Lab of Swine Genetics and Breeding, Ministry of Agriculture and Rural Affairs, Huazhong Agricultural University, Wuhan 430070, P.R. China; The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan 430070, P.R. China; Hubei Hongshan Laboratory, Frontiers Science Center for Animal Breeding and Sustainable Production, Wuhan 430070, P.R. China. Electronic address: xyli@mail.hzau.edu.cn.
Shuhong ZhaoKey Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education & Key Lab of Swine Genetics and Breeding, Ministry of Agriculture and Rural Affairs, Huazhong Agricultural University, Wuhan 430070, P.R. China; Yazhouwan National Laboratory, Sanya 572024, P.R. China; The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan 430070, P.R. China; Hubei Hongshan Laboratory, Frontiers Science Center for Animal Breeding and Sustainable Production, Wuhan 430070, P.R. China. Electronic address: shzhao@mail.hzau.edu.cn.
Shengsong XieKey Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education & Key Lab of Swine Genetics and Breeding, Ministry of Agriculture and Rural Affairs, Huazhong Agricultural University, Wuhan 430070, P.R. China; The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan 430070, P.R. China; Hubei Hongshan Laboratory, Frontiers Science Center for Animal Breeding and Sustainable Production, Wuhan 430070, P.R. China. Electronic address: ssxie@mail.hzau.edu.cn.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

CRISPR-Cas12a is a compact, RNA-guided nuclease widely deployed in genome editing and molecular diagnostics, yet its broader utility is limited by suboptimal cis-cleavage efficiency and incompletely defined trans-cleavage behavior. To overcome these constraints, we developed an artificial intelligence-guided structural discovery pipeline powered by AlphaFold2, which identified 1,261 previously uncharacterized Cas12a orthologs. From this set, 21 structurally conserved but sequence-divergent candidates were selected for biochemical characterization. Using structure-informed engineering, we generated PcuCas12a MAX, a high-fidelity variant that achieves genome-editing efficiencies in human cells comparable to the benchmark AsCas12a Ultra while retaining robust activity in murine and porcine systems. In addition, four orthologs (LcoCas12a, FcaCas12a, EsoCas12a, and Mac2Cas12a), when paired with specifically engineered CRISPR RNAs, exhibited distinct single-stranded DNA trans-cleavage signatures. These properties enabled construction of a multiplex CRISPR sensor capable of simultaneously detecting multiple nucleic acid targets. Together, these findings expand the Cas12a endonuclease repertoire and enhance its utility in genome engineering and next-generation diagnostics.

Indexed as

Bacterial ProteinsClustered Regularly Interspaced Short Palindromic RepeatsCRISPR-Associated ProteinsCRISPR-Cas SystemsEndodeoxyribonucleasesGene EditingMetagenomeAnimalsHumansMicePathology, MolecularRNA, Guide, CRISPR-Cas SystemsSwineBacterial ProteinsCas12a proteinCRISPR-Associated ProteinsEndodeoxyribonucleasesRNA, Guide, CRISPR-Cas SystemsCas12a orthologscis-/trans-cleavage activitygenome editingmultiplex diagnostics

Identifiers

PMID41814651
PMCPMC13330018

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.