Abstract
DNA/RNA-gold nanoparticle (DNA/RNA-AuNP) nanoprobes have been widely employed for nanobiotechnology applications. Here, we discover that both thiolated and non-thiolated DNA/RNA can be efficiently attached to AuNPs to achieve high-stable spherical nucleic acid (SNA) within minutes under a domestic microwave (MW)-assisted heating-dry circumstance. Further studies show that for non-thiolated DNA/RNA the conjugation is poly (T/U) tag dependent. Spectroscopy, test strip hybridization, and loading counting experiments indicate that low-affinity poly (T/U) tag mediates the formation of a standing-up conformation, which is distributed in the outer layer of SNA structure. In further application studies, CRISPR/Cas9-sgRNA (136 bp), SARS-CoV-2 RNA fragment (1278 bp), and rolling circle amplification (RCA) DNA products (over 1000 bp) can be successfully attached on AuNPs, which overcomes the routine methods in long-chain nucleic acid-AuNP conjugation, exhibiting great promise in biosensing and nucleic acids delivery applications. Current heating-dry strategy has improved traditional DNA/RNA-AuNP conjugation methods in simplicity, rapidity, cost, and universality.
Highlights
DNA/RNA-gold nanoparticle (DNA/RNA-AuNP) nanoprobes have been widely employed for nanobiotechnology applications
Spectroscopy, test strip hybridization, and loading counting experiments indicate that poly (T/U) tag is distributed in the outer layer of such a spherical nucleic acid (SNA) structure
Our method addresses the deficiencies of traditional methods in long DNA/RNA labeling and allows selective functionalization of AuNPs with sgRNA (136 bp), RNA (1278 bp) fragment encoding nucleocapsid (N) protein (GU280_gp[10] locus, termed N gene) of SARS-CoV-2, and rolling circle amplification (RCA) DNA products
Summary
DNA/RNA-gold nanoparticle (DNA/RNA-AuNP) nanoprobes have been widely employed for nanobiotechnology applications. Results showed that the number of poly (T)-tagged DNA probes attach to each AuNP by MW-assisted heating-dry method is 83, which is about 2 times higher than the salt-aging and low-pH method (Fig. 1f).
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