2026-10-10T04:22:50Z https://api.figshare.com/v2/oai

oai:figshare.com:article/33328818 2026-10-09T12:04:34Z category_25093 item_type_3 month_year_10_2026

<b>A Qualitative and Quantitative Detection Method for Gene Editor Residues Based on Cutting-Edge Biosensing Technology</b> Zizhao Wang (24647316) Sensor technology (incl. chemical aspects) Genetically modified organisms CRISPR/Cas technology Aptamer <p dir="ltr">This study focuses on the core pain points of the current gene editing industry, specifically addressing the residual editor issue arising from the widespread application of CRISPR/Cas9 technology. A novel, highly sensitive method for the qualitative and quantitative detection of residual Cas9 has been developed, which integrates CRISPR/Cas12a with aptamer-based recognition.By combining secondary structure prediction and tertiary structure molecular docking, the key binding sites of the Cas9-specific aptamer were validated. A fluorescence aptasensor based on a competitive binding mechanism was then constructed: in the presence of residual Cas9, the aptamer preferentially binds to the target, releasing complementary DNA (cDNA) that activates the trans-cleavage activity of Cas12a. This activity cleaves a single-stranded DNA (ssDNA) probe dual-labeled with a fluorophore and a quencher, generating a fluorescence signal (relative fluorescence units, RFU) that is positively correlated with the Cas9 concentration.Systematic optimization experiments determined the optimal detection conditions: a 20-minute trans-cleavage duration, a 1:1 molar ratio of Cas12a to CRISPR RNA (crRNA), a final aptamer concentration of 180 nM, and a combination of high- and low-salt buffers to balance molecular recognition specificity and protein stability.In conclusion, this study provides a low-cost, highly sensitive detection strategy, offering a novel technical pathway for the safety regulation of gene-edited products.</p> 2026-10-09T12:04:34Z Dataset Dataset 10.6084/m9.figshare.33328818.v3 https://figshare.com/articles/dataset/_b_A_Qualitative_and_Quantitative_Detection_Method_for_Gene_Editor_Residues_Based_on_Cutting-Edge_Biosensing_Technology_b_/33328818 CC BY 4.0