Cellular uptake of neutral phosphorodiamidate morpholino oligomers

Patrick L. Iversen, Katherine Aird, Rebecca Wu, Michael M. Morse, Gayathri R. Devi

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Phosphorodiamidate morpholino oligomers (PMO), which have a neutral chemistry, are extensively being used as tools for selective inhibition of gene expression in cell culture models and are currently in human clinical trials. Unlike phosphorothioates (PS ODN) and other charged oligonucleotides, little is known about the uptake characteristics of neutral oligomers. The purpose of this study was to understand the kinetics of PMO transport in cells and correlate with antisense activity. In contrast to primary cells and some transformed cell lines which were uptake permissive, established cancer cell lines showed very poor uptake with an occasional diffuse intracellular pattern. Differential PMO uptake was also observed in immune cells, with dendritic cells and monocytes showing highest uptake compared to T and B cells. In addition, PMO localization was observed to be heterogeneous within a population of uptake permissive cells. Unassisted PMO delivery targeting specific genes was correlated with functional antisense efficacy in experiments showing correction of pre-mRNA missplicing and inhibition of target enzyme activity in cells in culture. PMO internalization in uptakepermissive cells was identified to be specific, saturable, and energy-dependent, suggesting a receptor mediated uptake mechanism. Understanding PMO transport should facilitate the design of more effective synthetic antisense oligomers as therapeutic agents.

Original languageEnglish (US)
Pages (from-to)579-588
Number of pages10
JournalCurrent Pharmaceutical Biotechnology
Volume10
Issue number6
DOIs
StatePublished - Dec 16 2009

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Morpholinos
Cell Culture Techniques
Transformed Cell Line
Gene Targeting
RNA Precursors
Oligonucleotides
Dendritic Cells
Monocytes
B-Lymphocytes
Clinical Trials
T-Lymphocytes
Gene Expression
Cell Line
Enzymes
Population

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Pharmaceutical Science

Cite this

Iversen, Patrick L. ; Aird, Katherine ; Wu, Rebecca ; Morse, Michael M. ; Devi, Gayathri R. / Cellular uptake of neutral phosphorodiamidate morpholino oligomers. In: Current Pharmaceutical Biotechnology. 2009 ; Vol. 10, No. 6. pp. 579-588.
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Cellular uptake of neutral phosphorodiamidate morpholino oligomers. / Iversen, Patrick L.; Aird, Katherine; Wu, Rebecca; Morse, Michael M.; Devi, Gayathri R.

In: Current Pharmaceutical Biotechnology, Vol. 10, No. 6, 16.12.2009, p. 579-588.

Research output: Contribution to journalArticle

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