PMID- 15906394
OWN - NLM
STAT- MEDLINE
DCOM- 20051101
LR  - 20061115
IS  - 1040-452X (Print)
IS  - 1040-452X (Linking)
VI  - 72
IP  - 1
DP  - 2005 Sep
TI  - Multi-transgenic pigs expressing three fluorescent proteins produced with high 
      efficiency by sperm mediated gene transfer.
PG  - 68-76
AB  - 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.
FAU - Webster, Nicole L
AU  - Webster NL
AD  - The Austin Research Institute, Studley Road, Heidelberg, Victoria, Australia.
FAU - Forni, Monica
AU  - Forni M
FAU - Bacci, Maria Laura
AU  - Bacci ML
FAU - Giovannoni, Roberto
AU  - Giovannoni R
FAU - Razzini, Riccardo
AU  - Razzini R
FAU - Fantinati, Paolo
AU  - Fantinati P
FAU - Zannoni, Augusta
AU  - Zannoni A
FAU - Fusetti, Lisa
AU  - Fusetti L
FAU - Dalpra, Leda
AU  - Dalpra L
FAU - Bianco, Maria Rosaria
AU  - Bianco MR
FAU - Papa, Michele
AU  - Papa M
FAU - Seren, Eraldo
AU  - Seren E
FAU - Sandrin, Mauro S
AU  - Sandrin MS
FAU - Mc Kenzie, Ian F C
AU  - Mc Kenzie IF
FAU - Lavitrano, Marialuisa
AU  - Lavitrano M
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Mol Reprod Dev
JT  - Molecular reproduction and development
JID - 8903333
RN  - 147336-22-9 (Green Fluorescent Proteins)
SB  - IM
MH  - Animals
MH  - Animals, Genetically Modified/embryology/*genetics
MH  - Blastocyst/cytology/physiology
MH  - Female
MH  - Gene Expression Regulation, Developmental/genetics/*physiology
MH  - Gene Transfer Techniques
MH  - Green Fluorescent Proteins/*biosynthesis/genetics
MH  - Male
MH  - Mice
MH  - Morula/cytology/physiology
MH  - Spermatozoa/cytology/*metabolism
MH  - Swine/embryology/*genetics
EDAT- 2005/05/21 09:00
MHDA- 2005/11/03 09:00
CRDT- 2005/05/21 09:00
PHST- 2005/05/21 09:00 [pubmed]
PHST- 2005/11/03 09:00 [medline]
PHST- 2005/05/21 09:00 [entrez]
AID - 10.1002/mrd.20316 [doi]
PST - ppublish
SO  - Mol Reprod Dev. 2005 Sep;72(1):68-76. doi: 10.1002/mrd.20316.