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In vitro selection for a polymerase ribozyme with improved substrate binding
Author(s) -
Moretti Janina,
Huang TaYou,
Muller Ulrich
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.653.2
Subject(s) - ribozyme , ligase ribozyme , rna , polymerase , substrate (aquarium) , computational biology , hairpin ribozyme , biology , vs ribozyme , chemistry , dna , biophysics , biochemistry , microbiology and biotechnology , gene , ecology
The origin of life went through a stage in which RNAs acted both as genome and as catalyst—the RNA world. Re‐creating a self‐replicating RNA system in the lab would help us to understand what an RNA based world may have looked like and how it evolved into today's protein and DNA based world. A self‐replicating RNA system would require an RNA catalyst (ribozyme) capable of polymerizing RNA, to replicate the system. A ribozyme capable of catalyzing RNA polymerization has been developed, but its efficiency is about 100‐fold below the level required for self‐replication due to weak affinity for its substrate. To improve substrate binding we are using an in vitro selection approach. In this method the sequences of a few functional ribozymes are selected from a large pool (~ 10 14 ) of ribozyme variants. Previous selections covalently linked the ribozyme to its substrate thereby removing selective pressure for substrate binding affinity. A method recently published by the Unrau lab facilitates selection of polymerase ribozyme that are not covalently linked to their substrate by compartmentalizing ribozymes and substrates in emulsions with about 10 14 droplets. We have improved many aspects of this recently published method and will soon be able to perform the selection.