BE-CATCH: Bioamplifier-Equipped CRISPR-Cas12a Transduction System Coupled with Commercial Pregnancy Test Strips to Harness Signal-on Point-of-Care Detection DOI

Tongshan Zuo,

Xiang Chen, Yan Yu

et al.

Analytical Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: April 15, 2025

Repurposing existing commercial diagnostic equipment to enable portable analysis of diverse targets is driving the development affordable point-of-care testing (POCT). Interestingly, we found that goat antimouse IgG could replace human chorionic gonadotropin (hCG) make T line pregnancy test strips (PTS) appear red color and accordingly synthesized a novel signal output probe, which eliminated intricate hCG covalent coupling steps, meet multiple needs expanded POCT. Given this, introduced separation-free universal POCT strategy termed bioamplifier-equipped CRISPR-Cas12a transduction system coupled with PTS harness signal-on detection (BE-CATCH). Specifically, target inputs were converted amplified by multiplied strand displacement amplification-based bioamplifier, thereby activating Cas12a's trans-cleavage activity. Then, activated Cas12a would cleave connector indiscriminately, ultimately kept probe in free state; thus, be translated into colorimetric on PTS. This not only provided boosted sensitivity specificity but also enhanced user-friendliness maintaining mode. We demonstrated versatility BE-CATCH through selectively detecting miR-155 flap endonuclease 1. its broad adaptability, provide an appealing option broaden application biomedical diagnostics.

Language: Английский

CRISPR detection of cardiac tumor-associated microRNAs DOI

Youlin Fu,

Peng Zhang, Feng Chen

et al.

Molecular Biology Reports, Journal Year: 2025, Volume and Issue: 52(1)

Published: Jan. 11, 2025

Language: Английский

Citations

1
Boosting CRISPR/Cas12a intrinsic RNA detection capability through pseudo hybrid DNA-RNA substrate design DOI Creative Commons
Yi Liu, Jie Qiao, Junqi Zhang

et al.

Research Square (Research Square), Journal Year: 2025, Volume and Issue: unknown

Published: March 26, 2025

Abstract The CRISPR/Cas12a system is known for its intrinsic RNA-guided trans-cleavage activity; however, RNA detection sensitivity limited, with conventional methods typically achieving limits in the nanomolar range. Here, we report development of "Pseudo Hybrid DNA-RNA" (PHD) assay that significantly enhances capability Cas12a. PHD achieves a striking limit 7.7 pM using single crRNA and 33.8 fM pooled crRNAs. Importantly, this exhibits ultra-high specificity, capable distinguishing mutated target sequences at PAM-distal region. It can also detect ultrashort as short 6–8 nucleotides long RNAs complex secondary structures. Additionally, enables PAM-free attomolar-level DNA detection. We further demonstrate practical utility by successfully detecting miR-155 biomarkers HPV16 clinical samples. anticipate design principles established study be extended to other CRISPR/Cas enzymes, thereby accelerating powerful nucleic acid testing tools various applications.

Language: Английский

Citations

0
BE-CATCH: Bioamplifier-Equipped CRISPR-Cas12a Transduction System Coupled with Commercial Pregnancy Test Strips to Harness Signal-on Point-of-Care Detection DOI

Tongshan Zuo,

Xiang Chen, Yan Yu

et al.

Analytical Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: April 15, 2025

Repurposing existing commercial diagnostic equipment to enable portable analysis of diverse targets is driving the development affordable point-of-care testing (POCT). Interestingly, we found that goat antimouse IgG could replace human chorionic gonadotropin (hCG) make T line pregnancy test strips (PTS) appear red color and accordingly synthesized a novel signal output probe, which eliminated intricate hCG covalent coupling steps, meet multiple needs expanded POCT. Given this, introduced separation-free universal POCT strategy termed bioamplifier-equipped CRISPR-Cas12a transduction system coupled with PTS harness signal-on detection (BE-CATCH). Specifically, target inputs were converted amplified by multiplied strand displacement amplification-based bioamplifier, thereby activating Cas12a's trans-cleavage activity. Then, activated Cas12a would cleave connector indiscriminately, ultimately kept probe in free state; thus, be translated into colorimetric on PTS. This not only provided boosted sensitivity specificity but also enhanced user-friendliness maintaining mode. We demonstrated versatility BE-CATCH through selectively detecting miR-155 flap endonuclease 1. its broad adaptability, provide an appealing option broaden application biomedical diagnostics.

Language: Английский

Citations

0