2025 Hyper Recent •CC0 1.0 Universal

This work is dedicated to the public domain. No rights reserved.

Access Preprint From Server
June 29th, 2025
Version: 1
UT-Austin
biochemistry
biorxiv

One-Pot Isothermal Linear Amplification and Cas12a-based Nucleic Acid Detection

Sinan, S.Open in Google Scholar•Kooistra, R.Open in Google Scholar•Rajaraman, K.Open in Google Scholar•Islam, Z.Open in Google Scholar•Madan, D.Open in Google Scholar•Nalefski, E.Open in Google Scholar•Finkelstein, I. J.Open in Google Scholar

CRISPR-based nucleic acid diagnostics are a promising class of point-of-care tools that could dramatically improve healthcare outcomes for millions worldwide. However, these diagnostics require nucleic acid pre-amplification, an additional step that complicates deployment to low resource settings. Here, we developed CATNAP (Cas trans-nuclease detection of amplified products), a method that integrates isothermal linear DNA amplification with Cas12a detection in a single reaction. CATNAP uses a nicking enzyme and DNA polymerase to continuously generate single-stranded DNA, activating Cas12a\'s trans-cleavage activity without damaging the template. We optimized enzyme combinations, buffer conditions, and target selection to achieve high catalytic efficiency. CATNAP successfully distinguished between high- and low-risk HPV strains and detects HPV-16 in a cervical cancer crude cell lysate at room temperature with minimal equipment, offering advantages over PCR-based approaches. We conclude that CATNAP bridges the sensitivity gap in CRISPR diagnostics while maintaining simplicity, making accurate disease detection more accessible in resource-limited settings.

Similar Papers

biorxiv
Sun Jun 29 2025
Inhibition of Chikungunya virus nsP2 protease in vitro by pantinin-1 isolated from scorpion venom
Climate change has enhanced the spread of arboviruses such as Chikungunya virus (CHIKV). CHIKV is a re-emerging virus from the family Togaviridae that has spread globally, causing numerous outbreaks. The lack of antiviral therapy against CHIKV makes it a significant threat to public health. Cleavage of the viral polyprotein depends on the catalytic activity of nsP2, which is essential for viral re...
Mastalipour, M.
•
Coronado, M. A.
•
Gonzalez, J. E. H.
•
Willbold, D.
•
Eberle, R. J.
biorxiv
Sat Jun 28 2025
Stability and DNA Methyltransferase Activity of DNMT3A are Maintained by Ubiquitin-Specific Peptidase 11 (USP11) and Sumoylation Countering Degradation
DNA methyltransferase 3A (DNMT3A) plays crucial roles in mammalian development and hematopoiesis. DNMT3A protein instability is associated with blood diseases such as myelodysplastic syndrome (MDS), acute myeloid leukemia (AML), as well as Tatton-Brown-Rahman syndrome, an overgrowth disorder. We found that certain unstable DNMT3A mutations cause DNMT3A localization changes, resulting in loss of fu...
Yonezawa, T.
•
Rutter, J.
•
Ramabadran, R.
•
Sundaramurthy, V.
...•
Goodell, M. A.
biorxiv
Sat Jun 28 2025
Phosphoproteomics unveils the signaling dynamics in neuronal cells stimulated with insulin and insulin-like growth factors
Background: Given the role of metabolism in brain health and disease, investigating the role of insulin (INS) and insulin-like growth factors (IGFs) as potential therapeutic strategies for neurodegenerative diseases is currently underway. Yet, the signaling pathways associated with INS and IGFs in the brain remain elusive, particularly for the human brain. Unraveling these pathways is critical for...
Ushakumary, M. G.
•
Sontag, R.
•
Posso, C.
•
Fillmore, T. L.
...•
Petyuk, V. A.
biorxiv
Sat Jun 28 2025
Evaluating horseradish peroxidase-mimic DNAzyme transducer for glucometer readout: roles of various components and their optimization
The Development of decentralized disease detection elements is integral towards ensuring democratized access to healthcare. Personal glucometers, thanks to their low cost, ease of usage, and universal availability, are increasingly being identified as a central tenet of democratized disease detection. Towards this objective, it will first be critical to establish whether glucometers can be integra...
Parveen, S.
•
Ghosh, S.
•
Sengupta, M.
•
Talukdar, A.
biorxiv
Sat Jun 28 2025
Excretory-secretory products of the fish-borne parasite Anisakis simplex L3 larvae possess allergens and unusual glycan modifications
Anisakis simplex is a parasitic aquatic nematode, which may cause mild-to-severe gastrointestinal allergic reactions (Anisakiasis) with clinical symptoms, such as rhinitis and urticaria in humans who accidentally consume raw or undercooked marine products contaminated with infective L3 Anisakis larvae. Several Anisakis excretory/secretory (E/S) products and somatic proteins are known to be involve...
Adduci, I.
•
Wang, X.
•
Paschinger, K.
•
Wilson, I. B. H.
...•
Yan, S.
biorxiv
Sat Jun 28 2025
Influence of ULK1 activity on the memory of ATG101 auto-activation dynamics
The initiation of autophagy is marked by the prompt convergence of initiation proteins to form super molecular complexes in spatially-defined autophagic hubs. The recruitment and activation of the ULK-kinase complex is a pivotal event in starting the initiation cascade. ULK complex-component ATG101 is required for initiating autophagy in mammalian cells, but its function is unknown. Its HORMA doma...
Patel, A.
•
Schneider, S.
•
Nagy, G.
•
Dybkov, O.
...•
Faesen, A. C.
biorxiv
Sat Jun 28 2025
Unveiling the biochemical catalysis and regulatory mechanisms of the E3-independent E2 enzyme hUBE2O
UBE2O, an E3-independent E2 ubiquitin-conjugation enzyme, directly engages substrates to mediate Ubiquitin conjugation and ligation. Despite its critical role in ubiquitination of multiple substrates, the catalytic and regulatory mechanisms of Homo sapiens UBE2O (hUBE2O) remain incompletely understood. Here, combining domain truncation, systematic mutagenesis and well-designed biochemical approach...
Xiang, D.
•
Tang, X.
•
Shi, R.
•
Dong, S.
•
Zhang, X.
biorxiv
Sat Jun 28 2025
Novel mechanistic insights for catalytic bioluminescence of mammalian Gaussia Luciferase through mutant and ancestral analysis
A mechanistic basis for luciferase bioluminescence provides a glimpse into its evolutionary role for organism survival, as it provides a blueprint to engineer luciferase enzymes for advanced technological applications. Gaussia Luciferase is among the brightest natural luciferases, but (1) the evolutionary development of its luminescence behavior remains unclear, (2) recent fundamental studies util...
Borum, R. M.
•
Lanzillotti, M.
•
Sahasrabuddhe, A.
•
Hui, J.
...•
Ferbas, J.
biorxiv
Sat Jun 28 2025
Chemoenzymatic Synthesis of Asymmetric Bisecting Bi-, Tri-, and Tetra-Antennary N-Glycans
N-Acetylglucosaminyltransferase-III (GnT-III) is a glycosyltransferase that can install a {beta}1,4-linked N-acetylglucosamine (GlcNAc) residue at the central {beta}-mannoside of N-glycans. The resulting so-called bisecting GlcNAc is not further extended by glycosyl transferases and has been implicated a wide range of biological processes. The molecular mechanisms by which bisection modulates the ...
Ghotekar, B. K.
•
Bhagwat, S. K.
•
Chopra, P.
•
Buckley, T.
•
Boons, G.-J.