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Selection of targets and the most efficient hairpin ribozymes for inactivation of mRNAs using a self‐cleaving RNA library
Author(s) -
BarrosodelJesus Alicia,
BerzalHerranz Alfredo
Publication year - 2001
Publication title -
embo reports
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 4.584
H-Index - 184
eISSN - 1469-3178
pISSN - 1469-221X
DOI - 10.1093/embo-reports/kve247
Subject(s) - ribozyme , ligase ribozyme , computational biology , mammalian cpeb3 ribozyme , rna , biology , gene , vs ribozyme , selection (genetic algorithm) , hairpin ribozyme , identification (biology) , genetics , computer science , artificial intelligence , botany
The identification of proficient target sites within long RNA molecules, as well as the most efficient ribozymes for each, is a major concern for the use of ribozymes as gene suppressers. In vitro selection methods using combinatorial libraries are powerful tools for the rapid elucidation of interactions between macromolecules, and have been successfully used for different types of ribozyme study. This paper describes a new method for selecting effective target sites within long RNAs using a combinatorial library of self‐cleaving hairpin ribozymes that includes all possible specificities. The method also allows the identification of the most appropriate ribozyme for each identified site. Searching for targets within the lacZ gene with this strategy yielded a clearly accessible site. Sequence analysis of ribozymes identified two variants as the most appropriate for this site. Both selected ribozymes showed significant inhibitory activity in the cell milieu.