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Related Experiment Videos

Human adenovirus type 35: nucleotide sequence and vector development.

W Gao1, P D Robbins, A Gambotto

  • 1Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA 15219, USA.

Gene Therapy
|October 7, 2003
PubMed
Summary
This summary is machine-generated.

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Researchers sequenced the human adenovirus serotype 35 (Ad35) genome, identifying 47 open reading frames. Novel Ad35-based vectors were developed for gene therapy and vaccine applications, efficiently infecting dendritic cells.

Area of Science:

  • Virology
  • Genomics
  • Molecular Biology

Background:

  • Human adenovirus serotype 35 (Ad35) is a significant pathogen.
  • Understanding the Ad35 genome is crucial for developing novel therapeutic vectors.
  • Previous studies have characterized various adenovirus serotypes, but a complete genomic sequence for Ad35 was needed.

Purpose of the Study:

  • To determine the complete genomic sequence of the Ad35 Holden strain.
  • To identify and characterize open reading frames (ORFs) within the Ad35 genome.
  • To develop Ad35-based vector systems for gene therapy and vaccine development.

Main Methods:

  • Whole-genome sequencing of the Ad35 Holden strain.
  • Bioinformatic analysis to identify and compare open reading frames (ORFs).

Related Experiment Videos

  • Development of Ad35-based vectors with deletions in E1, E3, or both regions.
  • Establishment of an HEK293-derived cell line for vector propagation.
  • Testing of vector infectivity in human and rhesus macaque dendritic cells.

    • Main Results:

    • The complete 34,794 base pair genomic sequence of Ad35 Holden strain was elucidated.
    • 47 open reading frames (ORFs) were identified, including conserved early and late regions, along with 14 novel ORFs.
    • Ad35 exhibits high homology to other adenovirus serotypes like Ad7 and Ad3.
    • Novel E1-, E3-, and E1/E3-deleted Ad35-based vectors were successfully generated.
    • The E1-deleted Ad35 vector was efficiently produced in HEK293 cells and demonstrated effective infection of human and rhesus macaque dendritic cells.

    Conclusions:

    • The complete Ad35 genome sequence provides a foundation for understanding its biology and developing novel vectors.
    • Ad35-based vectors, particularly E1-deleted versions, are promising tools for gene therapy and DNA-based vaccine development.
    • The developed vectors and packaging cell lines offer a versatile platform for preclinical and clinical applications.