DNA dose and sequence dependence in sperm-mediated gene transfer

Ilaria Sciamanna, Simona Piccoli, Laura Barberi, Germana Zaccagnini, Anna Rosa Magnano, Roberto Giordano, Paolo Campedelli, Clague Hodgson, Rodolfo Lorenzini, Corrado Spadafora

Research output: Contribution to journalArticlepeer-review

Abstract

We have tested three parameters in sperm-mediated gene transfer assays with mice and pigs: (i) the epididymal versus ejaculated origin of sperm cells, (ii) the primary structure, and (iii) the amount of the challenging foreign DNA. We have found that the pVLCNhGH construct, of retrotransposon origin, causes a massive embryo lethality and yet increases the yield of genetic transformation among born animals of both species compared to viral constructs. Arrest of embryonic development is a DNA dosedependent effect, which is observed with high DNA doses, while lower doses are compatible with development. Finally, the overall efficiency of sperm-mediated gene transfer is higher when ejaculated, versus epididymal, spermatozoa are used. We suggest that this difference is related to the highly efficient apoptotic response in epididymal compared to ejaculated spermatozoa, triggered by the interaction of exogenous DNA molecules with the sperm membrane. (C) 2000 Wiley-Liss, Inc.

Original languageEnglish
Pages (from-to)301-305
Number of pages5
JournalMolecular Reproduction and Development
Volume56
Issue number2 SUPPL.
Publication statusPublished - 2000

Keywords

  • Fertilization
  • Recombinant DNA
  • Spermatozoa
  • Transgenic animals

ASJC Scopus subject areas

  • Genetics
  • Developmental Biology
  • Cell Biology

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