Premium
DNA dose and sequence dependence in sperm‐mediated gene transfer
Author(s) -
Sciamanna Ilaria,
Piccoli Simona,
Barberi Laura,
Zaccagnini Germana,
Magnano Anna Rosa,
Giordano Roberto,
Campedelli Paolo,
Hodgson Clague,
Lorenzini Rodolfo,
Spadafora Corrado
Publication year - 2000
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/(sici)1098-2795(200006)56:2+<301::aid-mrd20>3.0.co;2-b
Subject(s) - biology , exogenous dna , sperm , retrotransposon , zygote , gene , dna , embryo , andrology , semen , genetics , microbiology and biotechnology , embryogenesis , genome , transposable element , medicine
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 dose‐dependent 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. Mol. Reprod. Dev. 56:301–305, 2000. © 2000 Wiley‐Liss, Inc.