Zeptomole detection of a viral nucleic acid using a target-activated ribozyme
Open accessAbstract
We describe a strategy for the ultra-sensitive detection of nucleic acids using “half” ribozymes that are devoid of catalytic activity unless completed by a trans-acting target nucleic acid. The half-ribozyme concept was initially demonstrated using a construct derived from a multiple turnover Class I ligase. Iterative RNA selection was carried out to evolve this half-ribozyme into one activated by a conserved sequence present in the hepatitis C virus (HCV) genome. Following sequence optimization of substrate RNAs, this HCV-activated half-ribozyme displayed a maximal turnover rate of 69 min−1 (pH 8.3) and was induced in rate by approximately 2.6 × 10 9-fold by the HCV target. It detected the HCV target oligonucleotide in the zeptomole range (6700 molecules), a sensitivity of detection roughly 2.6 × 10 6-fold greater than that previously demonstrated by oligonucleotide-activated ribozymes, and one that is sufficient for molecular diagnostic applications.
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The class I ligase and some of its polymerase descendants can be divided into fragments that self-assemble and retain function.
“Examples of these constructs include the class I ligase and some of its polymerase descendants, which can be divided into fragments that self-assemble and retain function (22–24).”
From An RNA polymerase ribozyme that synthesizes its own ancestor (2020) · cites this paper as [22]
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- Authors
- VAISH, N. K., JADHAV, V. R., KOSSEN, K., PASKO, C., ANDREWS, L. E., MCSWIGGEN, J. A., POLISKY, B., & SEIWERT, S. D.
- ORCID
- JADHAV, VASANT R.
- Venue
- RNA
- Publisher
- Cold Spring Harbor Laboratory
- Published
- 2003-08-15
- Volume
- 9
- Issue
- 9
- Pages
- 1058-1072
- ISSN
- 1355-8382 (ISSN-L)
- 1469-9001
- PMC
- PMC1370471
- OpenAlex
- W1981113599
- Cited by
- 30
- References
- 50
- Citations by year
- 2026: 1 · 2022: 1 · 2020: 2 · 2018: 1 · 2015: 1 · 2012: 2
- FWCI
- 1.61
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- RNA (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Advanced biosensing and bioanalysis techniques
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
- Keywords
- zeptomole detection, hepatitis C virus, oligonucleotide detection, molecular diagnosis
- MeSH terms
- Hydrogen-Ion Concentration, RNA, Viral, Time Factors, Hepacivirus, RNA, Catalytic
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