Evidence map›Paper›PMID 34413466›Full record

ArticleCommunications biology2021

Protein or ribonucleoprotein-mediated blocking of recombinase polymerase amplification enables the discrimination of nucleotide and epigenetic differences between cell populations.

Toshitsugu Fujita, Shoko Nagata, Hodaka Fujii

Abstract read
In one paragraph

Article in Communications biology, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.

0numbers the graph read from it
0cells of the map it votes in
5citing 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

5 citing papers in PubMed.

  1. Droplet Digital CRISPR for Nucleic Acid Detection.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026
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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

3 authors.

Toshitsugu FujitaDepartment of Biochemistry and Genome Biology, Hirosaki University Graduate School of Medicine, Aomori, Japan. toshitsugu.fujita@hirosaki-u.ac.jp.ORCID 0000-0002-9536-9999
Shoko NagataDepartment of Biochemistry and Genome Biology, Hirosaki University Graduate School of Medicine, Aomori, Japan.
Hodaka FujiiDepartment of Biochemistry and Genome Biology, Hirosaki University Graduate School of Medicine, Aomori, Japan. hodaka@hirosaki-u.ac.jp.ORCID 0000-0003-1296-4256

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Isothermal DNA amplification, such as recombinase polymerase amplification (RPA), is well suited for point-of-care testing (POCT) as it does not require lengthy thermal cycling. By exploiting DNA amplification at low temperatures that do not denature heat-sensitive molecules such as proteins, we have developed a blocking RPA method to detect gene mutations and examine the epigenetic status of DNA. We found that both nucleic acid blockers and nuclease-dead clustered regularly interspaced short palindromic repeats (CRISPR) ribonucleoproteins suppress RPA reactions by blocking elongation by DNA polymerases in a sequence-specific manner. By examining these suppression events, we are able to discriminate single-nucleotide mutations in cancer cells and evaluate genome-editing events. Methyl-CpG binding proteins similarly inhibit elongation by DNA polymerases on CpG-methylated template DNA in our RPA reactions, allowing for the detection of methylated CpG islands. Thus, the use of heat-sensitive molecules such as proteins and ribonucleoprotein complexes as blockers in low-temperature isothermal DNA amplification reactions markedly expands the utility and application of these methods.

Indexed as

Clustered Regularly Interspaced Short Palindromic RepeatsEpigenesis, GeneticMutationCold TemperatureDNANucleic Acid Amplification TechniquesPoint-of-Care TestingRecombinasesRibonucleoproteinsDNARecombinasesRibonucleoproteins

Identifiers

PMID34413466
PMCPMC8376914

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LicenceCC BY
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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.