Document Detail

Efficient production of transgenic cloned calves using preimplantation screening.
MedLine Citation:
PMID:  12390880     Owner:  NLM     Status:  MEDLINE    
The genetic manipulation of donor cells before nuclear transfer (NT) enables prior selection for transgene integration. However, selection for genetically modified cells using antibiotic drugs often results in mixed populations, resulting in a mixture of transgenic and nontransgenic donor cells for NT. In this study, we attempted to develop efficient strategies for the generation of human bile salt-stimulated lipase (BSSL) transgenic cows. Preimplantation screening by either biopsy or green fluorescent protein (GFP) expression was used to detect NT-derived BSSL transgenic embryos to ensure that the calf born would be transgenic. We compared the development rates of NT-derived embryos from G418- and GFP-selected donor cells. There were no significant differences (P < 0.001) in cleavage rate (67.2% vs. 60.0%) and blastocyst formation rate (44.9% vs. 41.2%). We also compared the pregnancy rates of the G418/biopsy and GFP preimplantation screened NT-derived blastocysts. The Day 40 pregnancy rate of the G418/biopsy group (40%) was lower than that of the GFP group (57%), but the calf birth rate of the G418/biopsy group (40%) was higher than that of the GFP group (21%). Healthy BSSL transgenic calves were born after both screening processes. This is the first report of biopsy-screened cloned transgenic animals. The results suggest that both selection methods are useful for detecting transgenic NT embryos without negatively affecting their development into viable transgenic offspring.
Shu-Hung Chen; Todd D Vaught; Jeff A Monahan; Jeremy Boone; Elizabeth Emslie; Peter M Jobst; Ashley E Lamborn; Angelika Schnieke; Laura Robertson; Alan Colman; Yifan Dai; Irina A Polejaeva; David L Ayares
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Biology of reproduction     Volume:  67     ISSN:  0006-3363     ISO Abbreviation:  Biol. Reprod.     Publication Date:  2002 Nov 
Date Detail:
Created Date:  2002-10-22     Completed Date:  2003-05-21     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  0207224     Medline TA:  Biol Reprod     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1488-92     Citation Subset:  IM    
PPL Therapeutics Inc., Blacksburg, Virginia 24060, USA.
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MeSH Terms
Animals, Genetically Modified*
Animals, Newborn
Blastocyst / physiology*
Gene Dosage
Genes, Reporter
Genetic Testing / methods*
Green Fluorescent Proteins
Luminescent Proteins / genetics
Pregnancy Outcome
Pregnancy, Animal
Reproductive Techniques, Assisted
Sterol Esterase / biosynthesis,  genetics*
Reg. No./Substance:
0/Luminescent Proteins; 147336-22-9/Green Fluorescent Proteins; EC 3.1.1.-/bile salt-stimulated lipase; EC Esterase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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