CRISPR/Cas12a Corona Nanomachine for Detecting Circulating Tumor Nucleic Acids in Serum.

Anal Chem

State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China.

Published: December 2024

AI Article Synopsis

  • Circulating tumor nucleic acids (CTNAs) like cell-free DNA and RNA from tumors are potential biomarkers for cancer diagnosis and prognosis, highlighting the need for sensitive detection methods.
  • The study developed a CRISPR/Cas12a nanomachine that can detect tumor DNA and RNA effectively in serum, thanks to a protective shell that prevents breakdown by enzymes.
  • This innovative tool demonstrated the ability to simultaneously identify multiple types of CTNAs at very low concentrations, making it a promising method for early cancer diagnosis.

Article Abstract

Circulating tumor nucleic acids (CTNAs), which consist of cell-free DNA or RNA released from tumor cells, are utilized as potential biomarkers for diagnosing and managing tumor prognosis. There is a significant demand for developing a highly sensitive and reliable assay for CTNAs detection. In this study, we engineered a CRISPR/Cas12a corona nanomachine capable of detecting circulating tumor DNA and RNA in serum. This nanomachine consists of a protein shell incorporating Cas12a/crRNA ribonucleoprotein complexes and a scaffold AuNP core decorated with substrate ssDNA strands. The protective CRISPR corona shields the nucleic acid core from degradation by nuclease DNase/RNase, thereby enhancing the stability of the CRISPR/Cas12a corona nanomachine in biological fluids, even tolerating up to undiluted human serum and FBS. Upon encountering target CTNAs, the CRISPR/Cas12a is activated through the sequence-specific hybridization between crRNA and CTNAs. Subsequently, the activated CRISPR/Cas12a autonomously cleaves the collateral ssDNA substrates on AuNPs, releasing the fluorophore-labeled fragment and generating an increasing fluorescent signal. The CRISPR corona nanomachine was successfully employed to detect various CTNAs, including circulating tumor (ct)DNA/RNA ( L858R) and microRNA-21, achieving a limit of detection of 0.14 pM for ctDNA and 1.0 pM for RNA. This CRISPR corona nanomachine enables simultaneous detection of both DNA and RNA in complex biological samples, offering a promising tool for early diagnosis.

Download full-text PDF

Source
http://dx.doi.org/10.1021/acs.analchem.4c04993DOI Listing

Publication Analysis

Top Keywords

corona nanomachine
20
circulating tumor
16
crispr/cas12a corona
12
dna rna
12
crispr corona
12
detecting circulating
8
tumor nucleic
8
nucleic acids
8
nanomachine
6
tumor
6

Similar Publications

CRISPR/Cas12a Corona Nanomachine for Detecting Circulating Tumor Nucleic Acids in Serum.

Anal Chem

December 2024

State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China.

Article Synopsis
  • Circulating tumor nucleic acids (CTNAs) like cell-free DNA and RNA from tumors are potential biomarkers for cancer diagnosis and prognosis, highlighting the need for sensitive detection methods.
  • The study developed a CRISPR/Cas12a nanomachine that can detect tumor DNA and RNA effectively in serum, thanks to a protective shell that prevents breakdown by enzymes.
  • This innovative tool demonstrated the ability to simultaneously identify multiple types of CTNAs at very low concentrations, making it a promising method for early cancer diagnosis.
View Article and Find Full Text PDF

Near Infrared-Guided Smart Nanocarriers for MicroRNA-Controlled Release of Doxorubicin/siRNA with Intracellular ATP as Fuel.

ACS Nano

March 2016

State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, P. R. China.

In chemotherapy, it is a great challenge to recruit endogenous stimuli instead of external intervention for targeted delivery and controlled release; microRNAs are the most promising candidates due to their vital role during tumorigenesis and significant expression difference. Herein, to amplify the low abundant microRNAs in live cells, we designed a stimuli-responsive DNA Y-motif for codelivery of siRNA and Dox, in which the cargo release was achieved via enzyme-free cascade amplification with endogenous microRNA as trigger and ATP (or H(+)) as fuel through toehold-mediated strand displacement. Furthermore, to realize controlled release in tumor cells, smart nanocarriers were constructed with stimuli-responsive Y-motifs, gold nanorods, and temperature-sensitive polymers, whose surfaces could be reversibly switched between PEG and RGD states via photothermal conversion.

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!