Functional gene discovery using hybrid ribozyme libraries.
Hybrid ribozymes that couple the cleavage activity of hammerhead ribozymes with the unwinding activity of RNA helicases are powerful tools in the study of cell genetics and pharmaceutical drug development. They are useful for targeting a specific gene as well as screening functional genes to show phenotypic alterations. By randomizing the binding arms within the ribozymes, we can create a library of ribozymes that are capable of cleaving any mRNA. After introducing the library into cells and recovering the sequence information of ribozymes that alter the cell phenotype of interest, genes that are responsible for the specific phenotype can be identified. This chapter describes a method known as the gene-discovery system to identify novel functional genes related to a specific phenotype. Our gene-discovery system should be a powerful tool for post-genome research.
Kato
Yoshio
Y
Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, Japan.
Tsunemi
Masaru
M
Miyagishi
Makoto
M
Kawasaki
Hiroaki
H
Taira
Kazunari
K
D001483:Base Sequence; D017931:DNA Primers; D015723:Gene Library; D007202:Indicators and Reagents; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010641:Phenotype; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D055503:Protein Multimerization; D016337:RNA, Catalytic / Q000378:metabolism*; D015183:Restriction Mapping
D017931:DNA Primers; D007202:Indicators and Reagents; D016337:RNA, Catalytic; C427204:hammerhead ribozyme