Split Activator Assembly via Aptamer Binding Enables CRISPR-Cas12a Activation With Non-Nucleic Acid Targets.

Ram J Tharu, Yusha Imtiaz, Shubhajit Singha, Emmett Hanson, Reggie E Gold, Pavl Royzen, Mehmet V Yigit

Abstract read

In one paragraph

Article in Small methods, 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

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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

7 authors.

Ram J TharuDepartment of Chemistry, University At Albany, State University of New York, Albany, New York, USA.ORCID https://orcid.org/0009-0001-2402-1346

Yusha ImtiazDepartment of Chemistry, University At Albany, State University of New York, Albany, New York, USA.ORCID https://orcid.org/0000-0001-8997-6461

Shubhajit SinghaDepartment of Chemistry, University At Albany, State University of New York, Albany, New York, USA.ORCID https://orcid.org/0009-0003-2887-8480

Emmett HansonDepartment of Chemistry, University At Albany, State University of New York, Albany, New York, USA.ORCID https://orcid.org/0009-0000-9727-9682

Reggie E GoldDepartment of Chemistry, University At Albany, State University of New York, Albany, New York, USA.ORCID https://orcid.org/0009-0001-9836-5715

Pavl RoyzenDepartment of Chemistry, University At Albany, State University of New York, Albany, New York, USA.

Mehmet V YigitDepartment of Chemistry, University At Albany, State University of New York, Albany, New York, USA.ORCID https://orcid.org/0000-0002-4349-3701

Funding

DNA nanotechnology and synthetic biology for AI-supported detection and precision therapeuticsR35GM156250 · NIGMS · STATE UNIVERSITY OF NEW YORK AT ALBANY · PI Mehmet Veysel Yigit · 2025 to 2026

$779k

National Institute of General Medical Sciences of the National Institutes of Health R35GM156250NIGMS NIH HHS R35 GM156250

6 · The paper itself

Abstract

The programmability of CRISPR-Cas12a has enabled transformative advances in nucleic acid detection; however, extending its activation to non-nucleic-acid targets remains highly desirable. Here, we report a universal split-activator strategy that enables programmable Cas12a activation in response to a small molecule and a protein. In this approach, a canonical activator is divided into two fragments (p1 and p2), such that Cas12a activation occurs only upon their simultaneous assembly. We integrate this mechanism with aptamer-mediated conformation switching, wherein target binding destabilizes an aptamer-p2 duplex, releasing p2 to complete the activator complex and trigger Cas12a nuclease activity. Using this design, we demonstrate selective recognition of the small-molecule metabolite adenosine and the protein biomarker prostate-specific antigen (PSA), with high specificity against structurally related controls. Signal generation is achieved using fluorescent DNA nanoclusters (DFN-1), which undergo fluorescence decrease upon Cas12a-mediated degradation. The system exhibits concentration-dependent responses for both targets. Together, this work establishes a modular framework for coupling aptamer-based molecular recognition to programmable Cas12a activation, expanding the functional scope of CRISPR systems beyond nucleic acids and enabling new opportunities in CRISPR-Cas12a-mediated detection.

Indexed as

aptamerbiosensingCas12aDNA nanoclustersfluorescencesplit activator

Identifiers

PMID42806622

DOI10.1002/smtd.71075

PMCPMC13685422

What OpenQuestion holds

Textmetadata

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.