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A POCT assay based on commercial HCG strip for miRNA21 detection by integrating with RCA-HCR cascade amplification and CRISPR/Cas12a.
Mikrochim Acta
January 2025
Beijing Key Laboratory for Separation and Analysis in Biomedicine and Pharmaceuticals, School of Medical Technology, Beijing Institute of Technology, Beijing, 100081, China.
A point-of-care testing (POCT) assay based on commercial HCG strip was proposed for miRNA21 detection by integrating RCA-HCR cascaded isothermal amplification with CRISPR/Cas12a. Three modules were integrated in the proposed platform: target amplification module composed of rolling circle amplification (RCA) cascaded with hybridization chain reaction (HCR), signal transduction module composed of CRISPR/Cas12a combined with HCG-agarose gel beads probes, and signal readout module composed of commercial HCG strips. The proposed RCA-HCR-CRISPR/Cas12a-HCG strip assay for miRNA21 detection had high sensitivity, and the limit of detection was as low as 37 fM.
View Article and Find Full Text PDFNanopore-based consensus sequencing enables accurate multimodal tumor cell-free DNA profiling.
Genome Res
January 2025
University Medical Center Utrecht, Utrecht University, Oncode Institute, Cyclomics
Shallow genome-wide cell-free DNA (cfDNA) sequencing holds great promise for non-invasive cancer monitoring by providing reliable copy number alteration (CNA) and fragmentomic profiles. Single nucleotide variations (SNVs) are, however, much harder to identify with low sequencing depth due to sequencing errors. Here we present Nanopore Rolling Circle Amplification (RCA)-enhanced Consensus Sequencing (NanoRCS), which leverages RCA and consensus calling based on genome-wide long-read nanopore sequencing to enable simultaneous multimodal tumor fraction estimation through SNVs, CNAs, and fragmentomics.
View Article and Find Full Text PDFReplisomal coupling between the α-Pol III core and the τ-subunit of the clamp loader complex (CLC) are essential for genomic integrity in E. coli.
J Biol Chem
January 2025
Department of Chemistry and Biochemistry, Baylor University, Waco, Texas, 76798-7348, USA. Electronic address:
Coupling interactions between the alpha (α) subunit of the polymerase III core (α-Pol III core) and the tau (τ) subunit of the clamp loader complex (τ-CLC) are vital for efficient and rapid DNA replication in Escherichia coli (E. coli). Specific and targeted mutations in the C-terminal τ-interaction region of the Pol III α-subunit disrupted efficient coupled rolling circle DNA synthesis in vitro and caused significant genomic defects in CRISPR-Cas9 dnaE edited cell strains.
View Article and Find Full Text PDFTdT combined with Cas14a for the electrochemical biosensing of NPC-derived exosomes.
Bioelectrochemistry
January 2025
Shenzhen Baoan Authentic TCM Therapy Hospital, Shenzhen, 518101, China. Electronic address:
In this work, the electrochemical biosensor based on the subtle combination of terminal deoxynucleotidyl transferase (TdT), CRISPR/Cas14a, and magnetic nanoparticles (MNPs) was developed for the detection of nasopharyngeal carcinoma (NPC)-derived exosomes. Due to the synergistic effect of the following factors: the powerful elongation capacity of TdT for single-stranded DNA (ssDNA) with 3-hydroxy terminus, the outstanding trans-cleavage ability of CRISPR/Cas14a specifcally activated by the crRNA binding to target DNA, and the excellent separation ability of MNPs, the developed electrochemical biosensor exhibited high sensitivity for the detection of NPC-derived exosome, with a linear range from 6.0 × 10 ∼ 1.
View Article and Find Full Text PDFThe Circle Pure Rolling Method for Point Cloud Boundary Extraction.
Sensors (Basel)
December 2024
Facility Design and Instrumentation Institute, China Aerodynamics Research and Development Center, Mianyang 621000, China.
We introduce a circle rolling method (CRM) for boundary extraction from 2D point clouds. The core idea is to create a circle that performs pure rolling on the perimeter of the point cloud to obtain the boundary. For a 3D point cloud, a plane adsorbs points on both sides to create a 2D point cloud, and the CRM is used to extract the boundary points and map them back into space to obtain 3D boundary points.
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