Lisa Fusetti

Publications (2)5.06 Total impact

• Source

  Available from: Roberto Giovannoni

  Article: Multi-transgenic pigs expressing three fluorescent proteins produced with high efficiency by sperm mediated gene transfer.

  Nicole L Webster, Monica Forni, Maria Laura Bacci, Roberto Giovannoni, Riccardo Razzini, Paolo Fantinati, Augusta Zannoni, Lisa Fusetti, Leda Dalprà, Maria Rosaria Bianco, Michele Papa, Eraldo Seren, Mauro S Sandrin, Ian F C Mc Kenzie, Marialuisa Lavitrano
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  ABSTRACT: Multi-gene transgenic pigs would be of benefit for large animal models in medical, agricultural, and pharmaceutical applications; in particular for xenotransplantation, where extensive genetic manipulation of donor pigs is required to make them suitable for organ grafting to humans. We used the sperm mediated gene transfer (SMGT) method to produce with high efficiency multi-gene transgenic pigs using three genes coding for fluorescent proteins: enhanced blue (EBFP), green (EGFP), and red (DsRed2). All three fluorescent proteins were expressed in 171 out of 195 normally developed morula/blastocysts examined at day 6 post insemination (88%). Genomic DNA of 18 piglets born from two litters was screened by PCR, showing that all piglets were transgenic with at least one gene, 7/18 piglets were triple transgenic, 7/18 double transgenic, and 4/18 single transgenic. Fluorescence in situ hybridization (FISH) analysis revealed multiple sites of integration of the transgenes. RNA and protein expression was found in muscle, heart, liver, hair, and peripheral blood mononuclear cells (PBMCs). These results show that SMGT is an effective method for introducing multiple genes into pigs as shown by the simultaneous expression of three fluorescent proteins.
  Molecular Reproduction and Development 10/2005; 72(1):68-76. · 2.53 Impact Factor
• Article: Multi‐transgenic pigs expressing three fluorescent proteins produced with high efficiency by sperm mediated gene transfer

  Nicole L. Webster, Monica Forni, Maria Laura Bacci, Roberto Giovannoni, Riccardo Razzini, Paolo Fantinati, Augusta Zannoni, Lisa Fusetti, Leda Dalprà, Maria Rosaria Bianco, Michele Papa, Eraldo Seren, Mauro S. Sandrin, Ian F.C. Mc Kenzie, Marialuisa Lavitrano
  [show abstract] [hide abstract]
  ABSTRACT: Multi-gene transgenic pigs would be of benefit for large animal models in medical, agricultural, and pharmaceutical applications; in particular for xenotransplantation, where extensive genetic manipulation of donor pigs is required to make them suitable for organ grafting to humans. We used the sperm mediated gene transfer (SMGT) method to produce with high efficiency multi-gene transgenic pigs using three genes coding for fluorescent proteins: enhanced blue (EBFP), green (EGFP), and red (DsRed2). All three fluorescent proteins were expressed in 171 out of 195 normally developed morula/blastocysts examined at day 6 post insemination (88%). Genomic DNA of 18 piglets born from two litters was screened by PCR, showing that all piglets were transgenic with at least one gene, 7/18 piglets were triple transgenic, 7/18 double transgenic, and 4/18 single transgenic. Fluorescence in situ hybridization (FISH) analysis revealed multiple sites of integration of the transgenes. RNA and protein expression was found in muscle, heart, liver, hair, and peripheral blood mononuclear cells (PBMCs). These results show that SMGT is an effective method for introducing multiple genes into pigs as shown by the simultaneous expression of three fluorescent proteins. Mol. Reprod. Dev. © 2005 Wiley-Liss, Inc.
  Molecular Reproduction and Development 08/2005; 72(1):68 - 76. · 2.53 Impact Factor