Evidence map›Paper›PMID 37069831›Full record

ArticlePlant biotechnology journal2023

The engineered CRISPR-Mb2Cas12a variant enables sensitive and fast nucleic acid-based pathogens diagnostics in the field.

Jian Jiao, Yiqi Liu, Mengli Yang, Jingcheng Zheng, Chonghuai Liu, Wenxiu Ye, Shangwei Song, Tuanhui Bai, Chunhui Song, Miaomiao Wang and 6 more

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Plant biotechnology journal, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.

0numbers the graph read from it
0cells of the map it votes in
12citing 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

12 citing papers in PubMed.

  1. Article
  2. Molecular mechanisms and biotechnology applications of CRISPR-Cas12a.Nature reviews. Molecular cell biology · 2026
    Review
  3. Article
  4. Review
  5. Article
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  7. Article
  8. Review
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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

16 authors.

Jian JiaoCollege of Horticulture, Henan Agricultural University, Zhengzhou, China.ORCID 0000-0002-3947-6626
Yiqi LiuCollege of Horticulture, Henan Agricultural University, Zhengzhou, China.
Mengli YangCollege of Horticulture, Henan Agricultural University, Zhengzhou, China.
Jingcheng ZhengCollege of Horticulture, Henan Agricultural University, Zhengzhou, China.
Chonghuai LiuZhengzhou Fruit Research Institute, Chinese Academy of Agricultural Sciences, Zhengzhou, China.ORCID 0000-0001-6517-566X
Wenxiu YePeking University Institute of Advanced Agricultural Sciences, Shandong Laboratory of Advanced Agricultural Sciences at Weifang, Weifang, Shandong, China.ORCID 0000-0003-3464-507X
Shangwei SongCollege of Horticulture, Henan Agricultural University, Zhengzhou, China.
Tuanhui BaiCollege of Horticulture, Henan Agricultural University, Zhengzhou, China.
Chunhui SongCollege of Horticulture, Henan Agricultural University, Zhengzhou, China.
Miaomiao WangCollege of Horticulture, Henan Agricultural University, Zhengzhou, China.
Jiangli ShiCollege of Horticulture, Henan Agricultural University, Zhengzhou, China.
Ran WanCollege of Horticulture, Henan Agricultural University, Zhengzhou, China.
Kunxi ZhangCollege of Horticulture, Henan Agricultural University, Zhengzhou, China.
Pengbo HaoCollege of Horticulture, Henan Agricultural University, Zhengzhou, China.
Jiancan FengCollege of Horticulture, Henan Agricultural University, Zhengzhou, China.ORCID 0000-0001-7468-8491
Xianbo ZhengCollege of Horticulture, Henan Agricultural University, Zhengzhou, China.ORCID 0000-0001-7781-7594

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Existing CRISPR/Cas12a-based diagnostic platforms offer accurate and vigorous monitoring of nucleic acid targets, but have the potential to be further optimized for more efficient detection. Here, we profiled 16 Cas12a orthologs, focusing on their trans-cleavage activity and their potential as diagnostic enzymes. We observed the Mb2Cas12a has more robust trans-cleavage activity than other orthologs, especially at lower temperatures. An engineered Mb2Cas12a-RRVRR variant presented robust trans-cleavage activity and looser PAM constraints. Moreover, we found the existing one-pot assay, which simultaneously performed Recombinase Polymerase Amplification (RPA) and Cas12a reaction in one system, resulted in the loss of single-base discrimination during diagnosis. Therefore, we designed a reaction vessel that physically separated the RPA and Cas12a steps while maintaining a closed system. This isolated but closed system made diagnostics more sensitive and specific and effectively prevented contamination. This shelved Mb2Cas12a-RRVRR variant-mediated assay detected various targets in less than 15 min and exhibited equal or greater sensitivity than qPCR when detecting bacterial pathogens, plant RNA viruses and genetically modified crops. Overall, our findings further improved the efficiency of the current CRISPR-based diagnostic system and undoubtedly have great potential for highly sensitive and specific detection of multiple sample types.

Indexed as

Nucleic AcidsCRISPR-Cas SystemsCrops, AgriculturalPlants, Genetically ModifiedRecombinasesRNA, PlantNucleic AcidsRecombinasesRNA, PlantBacterial Cas12aCRISPR-Cas12aIn-field applicationNucleic acid detectionRPA

Identifiers

PMID37069831
PMCPMC10281610

What OpenQuestion holds

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LicenceCC BY-NC-ND
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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.