Design of a nonviral vector for site-selective, efficient integration into the human genome

Joseph Michael Kaminski, Mark R. Huber, James B. Summers, Matthew B. Ward

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Gene therapy in eukaryotes has met many obstacles. Research into the design of suitable nonviral vectors has been slow. To our knowledge, no nonviral vector has been proposed that allows for the possibility of highly efficient, site-selective integration into the genome of mammalian cells. On the basis of prior studies investigating the components necessary for transposon, retrovirus-like retrotransposon, and retroviral integration, we propose a nonviral system that would potentially allow for site-selective, efficient integration into the mammalian genome. Transposons have been developed that can transform a variety of cell lines. For example, the Sleeping Beauty transposon (SB) can transform a wide range of vertebrate cells from fish to human, and it mediates stable integration and long-term transgene expression in mice. However, the efficiency of transposition varies significantly among cell lines, suggesting the possible involvement of host factors in SB transposition. Here, we propose the use of a chimeric transposase (i.e., transposase-host DNA binding domain) to bypass the potential requirement of a host DNA-directing factor (or factors) for efficient, site-selective integration. We also discuss another potential method of docking the transposon-based vector adjacent to the host DNA, utilizing repetitive sequences for homologous recombination to promote efficient site-selective integration, as well as other site-selective nonviral approaches.

Original languageEnglish (US)
Pages (from-to)1242-1247
Number of pages6
JournalFASEB Journal
Volume16
Issue number10
DOIs
StatePublished - Aug 7 2002

Fingerprint

Human Genome
Transposases
Beauty
Genes
DNA
Cells
Genome
Cell Line
Retroelements
Nucleic Acid Repetitive Sequences
Homologous Recombination
Retroviridae
Eukaryota
Transgenes
Gene therapy
Genetic Therapy
Vertebrates
Fishes
Research Design
Fish

Keywords

  • Gene therapy
  • Recombination
  • Transposon

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

Cite this

Design of a nonviral vector for site-selective, efficient integration into the human genome. / Kaminski, Joseph Michael; Huber, Mark R.; Summers, James B.; Ward, Matthew B.

In: FASEB Journal, Vol. 16, No. 10, 07.08.2002, p. 1242-1247.

Research output: Contribution to journalArticle

Kaminski, Joseph Michael ; Huber, Mark R. ; Summers, James B. ; Ward, Matthew B. / Design of a nonviral vector for site-selective, efficient integration into the human genome. In: FASEB Journal. 2002 ; Vol. 16, No. 10. pp. 1242-1247.
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