Abstract：
:The Neurospora VS ribozyme is a catalytic RNA that has the unique ability to specifically recognize and cleave a stem-loop substrate through formation of a highly stable kissing-loop interaction (KLI). In order to explore the engineering potential of the VS ribozyme to cleave alternate substrates, we substituted the wild-type KLI by other known KLIs using an innovative engineering method that combines rational and combinatorial approaches. A bioinformatic search of the protein data bank was initially performed to identify KLIs that are structurally similar to the one found in the VS ribozyme. Next, substrate/ribozyme (S/R) pairs that incorporate these alternative KLIs were kinetically and structurally characterized. Interestingly, several of the resulting S/R pairs allowed substrate cleavage with substantial catalytic efficiency, although with reduced activity compared to the reference S/R pair. Overall, this study describes an innovative approach for RNA engineering and establishes that the KLI of the trans VS ribozyme can be adapted to cleave other folded RNA substrates.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Lacroix-Labonté J,Girard N,Dagenais P,Legault Pdoi
10.1093/nar/gkw401subject
Has Abstractpub_date
2016-08-19 00:00:00pages
6924-34issue
14eissn
0305-1048issn
1362-4962pii
gkw401journal_volume
44pub_type
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doi：10.1093/nar/gkh342
更新日期：2004-03-12 00:00:00
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journal_title：Nucleic acids research
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更新日期：2018-09-28 00:00:00
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pub_type： 杂志文章
doi：10.1093/nar/27.19.3899
更新日期：1999-10-01 00:00:00
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更新日期：1980-09-25 00:00:00
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更新日期：2017-05-19 00:00:00
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journal_title：Nucleic acids research
pub_type： 杂志文章
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更新日期：1987-12-23 00:00:00
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更新日期：2006-01-01 00:00:00
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journal_title：Nucleic acids research
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journal_title：Nucleic acids research
pub_type： 杂志文章
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