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Automated selection of aptamers against protein targets translated in vitro: from gene to aptamer
Open accessAbstract
Reagents for proteome research must of necessity be generated by high throughput methods. Aptamers are potentially useful as reagents to identify and quantitate individual proteins, yet are currently produced for the most part by manual selection procedures. We have developed automated selection methods, but must still individually purify protein targets. Therefore, we have attempted to select aptamers against protein targets generated by in vitro transcription and translation of individual genes. In order to specifically immobilize the protein targets for selection, they are also biotinylated in vitro. As a proof of this method, we have selected aptamers against translated human U1A, a component of the nuclear spliceosome. Selected sequences demonstrated exquisite mimicry of natural binding sequences and structures. These results not only reveal a potential path to the high throughput generation of aptamers, but also yield insights into the incredible specificity of the U1A protein for its natural RNA ligands.
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Mathews, D. H., Sabina, J., Zuker, M., & Turner, D. H. (1999) · Journal of Molecular Biology · doi:10.1006/jmbi.1999.2700 · Cited by 3,795
Cited by
Keefe, A. D., Pai, S., & Ellington, A. (2010) · Nature Reviews Drug Discovery · doi:10.1038/nrd3141 · Cited by 2,289
Synthetic biology: applications come of age
Khalil, A. S., & Collins, J. J. (2010) · Nature Reviews Genetics · doi:10.1038/nrg2775 · Cited by 1,373
Analytical applications of aptamers
Tombelli, S., Minunni, M., & Mascini, M. (2005) · Biosensors and Bioelectronics · doi:10.1016/j.bios.2004.11.006 · Cited by 1,017
Analysis of aptamer discovery and technology
Dunn, M. R., Jimenez, R. M., & Chaput, J. C. (2017) · Nature Reviews Chemistry · doi:10.1038/s41570-017-0076 · Cited by 938
Micromechanical Detection of Proteins Using Aptamer-Based Receptor Molecules
Savran, C. A., Knudsen, S. M., Ellington, A. D., & Manalis, S. R. (2004) · Analytical Chemistry · doi:10.1021/ac049859f · Cited by 310
Metadata
- Authors
- Cox, J. C.
- Venue
- Nucleic Acids Research
- Publisher
- Oxford University Press (OUP)
- Published
- 2002-10-15
- Volume
- 30
- Issue
- 20
- Pages
- 108e-108
- ISSN
- 0305-1048 (ISSN-L)
- 1362-4962
- OpenAlex
- W2132151485
- Cited by
- 173
- References
- 1
- FWCI
- 2.81
- Type
- journal article
- Language
- en
- Open access
- green
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