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  • 标题:SELEX-Based Screening of Exosome-Tropic RNA
  • 本地全文:下载
  • 作者:Takuma Yamashita ; Haruka Shinotsuka ; Yuki Takahashi
  • 期刊名称:Biological and Pharmaceutical Bulletin
  • 印刷版ISSN:0918-6158
  • 电子版ISSN:1347-5215
  • 出版年度:2017
  • 卷号:40
  • 期号:12
  • 页码:2140-2145
  • DOI:10.1248/bpb.b17-00519
  • 语种:English
  • 出版社:The Pharmaceutical Society of Japan
  • 摘要:

    Cell-derived nanosized vesicles or exosomes are expected to become delivery carriers for functional RNAs, such as small interfering RNA (siRNA). A method to efficiently load functional RNAs into exosomes is required for the development of exosome-based delivery carriers of functional RNAs. However, there is no method to find exosome-tropic exogenous RNA sequences. In this study, we used a systematic evolution of ligands by exponential enrichment (SELEX) method to screen exosome-tropic RNAs that can be used to load functional RNAs into exosomes by conjugation. Pooled single stranded 80-base RNAs, each of which contains a randomized 40-base sequence, were transfected into B16-BL6 murine melanoma cells and exosomes were collected from the cells. RNAs extracted from the exosomes were subjected to next round of SELEX. Cloning and sequencing of RNAs in SELEX-screened RNA pools showed that 29 of 56 clones had a typical RNA sequence. The sequence found by SELEX was enriched in exosomes after transfection to B16-BL6 cells. The results show that the SELEX-based method can be used for screening of exosome-tropic RNAs.

  • 关键词:exosome;aptamer;systematic evolution of ligands by exponential enrichment (SELEX)
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