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Optimizing aptamer activity for gene therapy applications using expression cassette SELEX.

Publication ,  Journal Article
Martell, RE; Nevins, JR; Sullenger, BA
Published in: Mol Ther
July 2002

RNA aptamers against a variety of clinically relevant target proteins have been generated. For example, we previously isolated an RNA aptamer that inhibits the function of the E2F family of transcription factors that play a critical role in the control of cell proliferation. However, the development of this and other aptamers for gene therapy applications has been complicated by the fact that expression of RNA aptamers in the context of flanking sequences can inhibit the ability of an aptamer to fold into its functional conformation. Insertion of the E2F aptamer into a tRNA expression cassette resulted in the production of high levels of chimeric tRNA that contains a misfolded and inactive aptamer in transfected mammalian cells. To overcome this problem, we randomized the sequence flanking the aptamer and selected for chimeric tRNAs that retained high affinity binding to E2F1. This expression cassette SELEX strategy yielded RNAs that bind E2F with high affinity (IC50 of 15 nM) and which can be expressed at high levels in mammalian cells. Moreover, these chimeric tRNA-E2F aptamers are functional and can inhibit E2F-mediated transactivation by up to 80% in human 293 cells. Expression cassette SELEX should greatly facilitate the use of aptamers for a variety of gene therapy applications.

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Published In

Mol Ther

DOI

ISSN

1525-0016

Publication Date

July 2002

Volume

6

Issue

1

Start / End Page

30 / 34

Location

United States

Related Subject Headings

  • Transcription Factors
  • RNA
  • Humans
  • Genetic Vectors
  • Genetic Therapy
  • E2F1 Transcription Factor
  • E2F Transcription Factors
  • DNA-Binding Proteins
  • DNA
  • Cell Cycle Proteins
 

Citation

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Martell, R. E., Nevins, J. R., & Sullenger, B. A. (2002). Optimizing aptamer activity for gene therapy applications using expression cassette SELEX. Mol Ther, 6(1), 30–34. https://doi.org/10.1006/mthe.2002.0624
Martell, Robert E., Joseph R. Nevins, and Bruce A. Sullenger. “Optimizing aptamer activity for gene therapy applications using expression cassette SELEX.Mol Ther 6, no. 1 (July 2002): 30–34. https://doi.org/10.1006/mthe.2002.0624.
Martell RE, Nevins JR, Sullenger BA. Optimizing aptamer activity for gene therapy applications using expression cassette SELEX. Mol Ther. 2002 Jul;6(1):30–4.
Martell, Robert E., et al. “Optimizing aptamer activity for gene therapy applications using expression cassette SELEX.Mol Ther, vol. 6, no. 1, July 2002, pp. 30–34. Pubmed, doi:10.1006/mthe.2002.0624.
Martell RE, Nevins JR, Sullenger BA. Optimizing aptamer activity for gene therapy applications using expression cassette SELEX. Mol Ther. 2002 Jul;6(1):30–34.
Journal cover image

Published In

Mol Ther

DOI

ISSN

1525-0016

Publication Date

July 2002

Volume

6

Issue

1

Start / End Page

30 / 34

Location

United States

Related Subject Headings

  • Transcription Factors
  • RNA
  • Humans
  • Genetic Vectors
  • Genetic Therapy
  • E2F1 Transcription Factor
  • E2F Transcription Factors
  • DNA-Binding Proteins
  • DNA
  • Cell Cycle Proteins