| Laser-mediated, site-specific inactivation of RNA transcripts. | |
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MedLine Citation:
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PMID: 10339553 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The biological function of specific gene products often is determined experimentally by blocking their expression in an organism and observing the resulting phenotype. Chromophore-assisted laser inactivation using malachite green (MG)-tagged antibodies makes it possible to inactivate target proteins in a highly restricted manner, probing their temporally and spatially resolved functions. In this report, we describe the isolation and in vitro characterization of a MG-binding RNA motif that may enable the same high-resolution analysis of gene function specifically at the RNA level (RNA-chromophore-assisted laser inactivation). A well-defined asymmetric internal bulge within an RNA duplex allows high affinity and high specificity binding by MG. Laser irradiation in the presence of low concentrations of MG induces destruction of the MG-binding RNA but not of coincubated control RNA. Laser-induced hydrolysis of the MG-binding RNA is restricted predominantly to a single nucleotide within the bulge. By appropriately incorporating this motif into a target gene, transcripts generated by the gene may be effectively tagged for laser-mediated destruction. |
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Authors:
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D Grate; C Wilson |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Proceedings of the National Academy of Sciences of the United States of America Volume: 96 ISSN: 0027-8424 ISO Abbreviation: Proc. Natl. Acad. Sci. U.S.A. Publication Date: 1999 May |
Date Detail:
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Created Date: 1999-06-24 Completed Date: 1999-06-24 Revised Date: 2013-04-17 |
Medline Journal Info:
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Nlm Unique ID: 7505876 Medline TA: Proc Natl Acad Sci U S A Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 6131-6 Citation Subset: IM |
Affiliation:
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Department of Biology and Center for the Molecular Biology of RNA, Sinsheimer Laboratories, University of California, Santa Cruz, CA 95064, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Antibodies Base Sequence Binding Sites Chromatography, Affinity DNA Primers Lasers* Molecular Sequence Data Nucleic Acid Conformation Polymerase Chain Reaction RNA / chemistry*, genetics*, radiation effects Rosaniline Dyes Sepharose Transcription, Genetic / radiation effects* |
| Chemical | |
Reg. No./Substance:
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0/Antibodies; 0/DNA Primers; 0/Rosaniline Dyes; 12058M7ORO/malachite green; 63231-63-0/RNA; 9012-36-6/Sepharose |
| Comments/Corrections | |
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