Isolation of a site-specifically modified RNA from an unmodified transcript

Natural RNAs contain many base modifications that have specific biological functions. The ability to functionally dissect individual modifications is facilitated by the identification and cloning of enzymes responsible for these modifications, but is hindered by the difficulty of isolating site-spec...

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Detalles Bibliográficos
Autores principales: Hou, Ya-Ming, Li, Zhi, Gamper, Howard
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2006
Materias:
Methods Online
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1363780/
https://www.ncbi.nlm.nih.gov/pubmed/16473844
http://dx.doi.org/10.1093/nar/gnj018
Descripción
Sumario:Natural RNAs contain many base modifications that have specific biological functions. The ability to functionally dissect individual modifications is facilitated by the identification and cloning of enzymes responsible for these modifications, but is hindered by the difficulty of isolating site-specifically modified RNAs away from unmodified transcripts. Using the m1G37 and m1A58 methyl modifications of tRNA as two examples, we demonstrate that non-pairing base modifications protect RNAs against the DNA-directed RNase H cleavage. This provide a new approach to obtain homogeneous RNAs with site-specific base modifications that are suitable for biochemical and functional studies.

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