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H1 RNA polymerase III promoter-driven expression of an RNA aptamer leads to high-level inhibition of intracellular protein activity.

Publication ,  Journal Article
Mi, J; Zhang, X; Rabbani, ZN; Liu, Y; Su, Z; Vujaskovic, Z; Kontos, CD; Sullenger, BA; Clary, BM
Published in: Nucleic Acids Res
2006

Aptamers offer advantages over other oligonucleotide-based approaches that artificially interfere with target gene function due to their ability to bind protein products of these genes with high affinity and specificity. However, RNA aptamers are limited in their ability to target intracellular proteins since even nuclease-resistant aptamers do not efficiently enter the intracellular compartments. Moreover, attempts at expressing RNA aptamers within mammalian cells through vector-based approaches have been hampered by the presence of additional flanking sequences in expressed RNA aptamers, which may alter their functional conformation. In this report, we successfully expressed a 'pure' RNA aptamer specific for NF-kappaB p50 protein (A-p50) utilizing an adenoviral vector employing the H1 RNA polymerase III promoter. Binding of the expressed aptamer to its target and subsequent inhibition of NF-kappaB mediated intracellular events were demonstrated in human lung adenocarcinoma cells (A549), murine mammary carcinoma cells (4T1) as well as a human tumor xenograft model. This success highlights the promise of RNA aptamers to effectively target intracellular proteins for in vitro discovery and in vivo applications.

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Published In

Nucleic Acids Res

DOI

EISSN

1362-4962

Publication Date

2006

Volume

34

Issue

12

Start / End Page

3577 / 3584

Location

England

Related Subject Headings

  • Xenograft Model Antitumor Assays
  • Transcriptional Activation
  • RNA Polymerase III
  • Promoter Regions, Genetic
  • Neoplasms
  • NF-kappa B p50 Subunit
  • Mice
  • Humans
  • Genetic Vectors
  • Gene Expression
 

Citation

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Mi, J., Zhang, X., Rabbani, Z. N., Liu, Y., Su, Z., Vujaskovic, Z., … Clary, B. M. (2006). H1 RNA polymerase III promoter-driven expression of an RNA aptamer leads to high-level inhibition of intracellular protein activity. Nucleic Acids Res, 34(12), 3577–3584. https://doi.org/10.1093/nar/gkl482
Mi, Jing, Xiuwu Zhang, Zahid N. Rabbani, Yingmiao Liu, Zhen Su, Zeljko Vujaskovic, Christopher D. Kontos, Bruce A. Sullenger, and Bryan M. Clary. “H1 RNA polymerase III promoter-driven expression of an RNA aptamer leads to high-level inhibition of intracellular protein activity.Nucleic Acids Res 34, no. 12 (2006): 3577–84. https://doi.org/10.1093/nar/gkl482.
Mi J, Zhang X, Rabbani ZN, Liu Y, Su Z, Vujaskovic Z, et al. H1 RNA polymerase III promoter-driven expression of an RNA aptamer leads to high-level inhibition of intracellular protein activity. Nucleic Acids Res. 2006;34(12):3577–84.
Mi, Jing, et al. “H1 RNA polymerase III promoter-driven expression of an RNA aptamer leads to high-level inhibition of intracellular protein activity.Nucleic Acids Res, vol. 34, no. 12, 2006, pp. 3577–84. Pubmed, doi:10.1093/nar/gkl482.
Mi J, Zhang X, Rabbani ZN, Liu Y, Su Z, Vujaskovic Z, Kontos CD, Sullenger BA, Clary BM. H1 RNA polymerase III promoter-driven expression of an RNA aptamer leads to high-level inhibition of intracellular protein activity. Nucleic Acids Res. 2006;34(12):3577–3584.
Journal cover image

Published In

Nucleic Acids Res

DOI

EISSN

1362-4962

Publication Date

2006

Volume

34

Issue

12

Start / End Page

3577 / 3584

Location

England

Related Subject Headings

  • Xenograft Model Antitumor Assays
  • Transcriptional Activation
  • RNA Polymerase III
  • Promoter Regions, Genetic
  • Neoplasms
  • NF-kappa B p50 Subunit
  • Mice
  • Humans
  • Genetic Vectors
  • Gene Expression