Enhancement of MSH receptor- and GAL4-mediated gene transfer by switching the nuclear import pathway

C. K. Chan, D. A. Jans

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Efficient nuclear delivery of plasmid DNA represents a major barrier in nonviral gene transfer. One approach has been to use DNA-binding proteins such as GAL4 from yeast as DNA carriers with nuclear targeting properties. We recently showed, however, that GAL4 is inefficient in targeting DNA to the nucleus because its DNA-binding and nuclear targeting activities are mutually exclusive, which relates to the fact that GAL4 nuclear import occurs via a novel pathway. Here, we 'switch' this pathway to a more conventional one by adding a modified poly-lysine to which an optimized nuclear targeting signal, based on that of the SV40 large T-antigen, is linked. We also use a chimeric GAL4-α-melanocyte stimulating hormone (MSH) fusion protein to enable gene transfer to cells expressing the MSH receptor. Switching the nuclear import pathway of the transfecting complex significantly enhances receptor-mediated gene transfer through enabling interaction with desired components of the cellular nuclear import machinery. The present study represents the first demonstration that nuclear targeting signals can enhance receptor-mediated gene delivery, the approaches having important relevance to research and clinical applications, such as in generating transgenic or knock-out animals, or in gene therapy.

Original languageEnglish
Pages (from-to)166-171
Number of pages6
JournalGene Therapy
Volume8
Issue number2
DOIs
Publication statusPublished - 2001
Externally publishedYes

Fingerprint

Cell Nucleus Active Transport
DNA
Genes
Melanocyte-Stimulating Hormones
Polyomavirus Transforming Antigens
Viral Tumor Antigens
DNA-Binding Proteins
Genetic Therapy
Lysine
Plasmids
Yeasts
MSH receptor
Research
Proteins

Keywords

  • GAL4 DNA-binding domain
  • Importin subunits
  • Melanocyte stimulating hormone (MSH)
  • Nuclear localization signals
  • Receptor-mediated gene transfer

ASJC Scopus subject areas

  • Genetics

Cite this

Enhancement of MSH receptor- and GAL4-mediated gene transfer by switching the nuclear import pathway. / Chan, C. K.; Jans, D. A.

In: Gene Therapy, Vol. 8, No. 2, 2001, p. 166-171.

Research output: Contribution to journalArticle

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