In vitro and in vivo production and purification of circular RNA aptamer

So Umekage, Yo Kikuchi

研究成果: Article

23 引用 (Scopus)

抜粋

RNA aptamers are potential candidates for RNA therapeutics. They must be clinically modified for medical applications because they are vulnerable to indigenous ribonucleases. Since circular RNA molecules without any chemical modification are much more stable than linear ones in a cell extract, we report the production of a circular form of streptavidin RNA aptamer both in vitro and in vivo. Circularization was accomplished by self-splicing permuted intron-exon sequences derived from T4 bacteriophage gene td. This sequence was producible in both Escherichia coli cells and in vitro. The circularized streptavidin RNA aptamer retained its binding of streptavidin and was stabile in HeLa cell extracts compared to the linear form of the streptavidin aptamer. The self-spliced circular RNA from the transcribed permuted intron-exon transcripts in E. coli cells was purified from a total RNA fraction using the solid-phase DNA probe method following anion exchange chromatography that excluded gel electrophoresis. This study provides an alternative method for designing and purifying useful RNA aptamers.

元の言語English
ページ(範囲)265-272
ページ数8
ジャーナルJournal of Biotechnology
139
発行部数4
DOI
出版物ステータスPublished - 2009 2 23
外部発表Yes

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

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