The stepwise endonuclease activity of a thermophilic Argonaute protein
Xun, G.; Liu, Q.; Chong, Y.; Li, Z.; Guo, X.; Li, Y.; Fei, H.; Li, K.; Feng, Y.
Show abstract
Thermophilic Argonaute proteins (Agos) can function as endonucleases via specific guide-target base-pairing cleavage for host defense. The ability to cleave target DNA sequences at any arbitrary sites endows them with reprogramed DNA capacity. Here, we identify that an Ago from the hyperthermophilic archaeon Pyrococcus furiosus (PfAgo) shows a stepwise endonuclease activity, which is demonstrated by the double strand DNA cleavage directed by a single guide DNA rather than canonical one pair of guide DNAs. We reveal that the cleavage products with 5-phosphorylated ends can used as the renewed guide which is capable to induce next round of cleavage to complementary sequences of target DNA. By combining the PfAgo stepwise endonuclease activity followed by target DNA amplification, we establish a rapid and specific platform for the unambiguously multiplex gene detection, termed RADAR (Renewed-gDNA Assisted DNA-cleavage by Argonaute). In the end, RADAR was applied to distinguish of human papillomavirus of serotypes in patient samples in a single reaction, suggesting that our technique would be adopted for diagnosing application.
Matching journals
The top 6 journals account for 50% of the predicted probability mass.
Similar papers in this journal
- Enhancement of Target Specificity of CRISPR-Cas12a by Using a Chimeric DNA-RNA Guide 94%
- Engineering an Escherichia coli strain for production of long single-stranded DNA 93%
- Constraints on the Emergence of RNA through Non-Templated Primer Extension with Mixtures of Potentially Prebiotic Nucleotides 93%
Similar papers in this journal
- CRISPR-Cas12a2-based rapid and sensitive detection system for target nucleic acid 93%
- Enzymatic Assembly for CRISPR Split-Cas9 System: The Emergence of a Sortase-based Split-Cas9 Technology 92%
- Selection of streptococcal glucan-binding protein C specific DNA aptamers to inhibit biofilm formation 92%
Similar papers in this journal
- All-in-One Dual CRISPR-Cas12a (AIOD-CRISPR) Assay: A Case for Rapid, Ultrasensitive and Visual Detection of Novel Coronavirus SARS-CoV-2 and HIV virus 96%
- Enhancement of trans-cleavage activity of Cas12a with engineered crRNA enables amplified nucleic acid detection 95%
- Comparative analysis of methodologies for detecting extrachromosomal circular DNA 94%
"Similar papers" are the closest papers from that journal in the model's embedding space. They show what the match is built on, but the ranking comes mostly from a classifier over the whole training set, not from these examples alone.