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Published on: July 30, 2020
Method of oocyte activation affects cloning efficiency in pigs.
Kristin M Whitworth1, Rongfeng Li, Lee D Spate
1Division of Animal Sciences, University of Missouri, Columbia, Missouri 65211, USA.
Molecular Reproduction and Development
|December 18, 2008
Summary
Comparing three somatic cell nuclear transfer (SCNT) protocols in pigs, the study found that using a proteasomal inhibitor (MG132) after fusion/activation significantly improved pregnancy rates, leading to live, genetically modified piglets.
Area of Science:
- Animal Biotechnology
- Reproductive Biology
- Developmental Biology
Background:
- Somatic cell nuclear transfer (SCNT) is crucial for producing genetically modified animals.
- Optimizing fusion and activation protocols is essential for improving SCNT efficiency in pigs.
- Porcine somatic cells expressing enhanced green fluorescent protein (pCAGG-EGFP) were used to evaluate different SCNT methods.
Purpose of the Study:
- To compare the efficiency of three distinct fusion/activation protocols for porcine SCNT.
- To assess the impact of a proteasomal inhibitor (MG132) on SCNT outcomes.
- To determine the optimal protocol for generating live, genetically modified piglets.
Main Methods:
- Three protocols were tested: electrical fusion/activation (NT1), electrical fusion/activation with MG132 (NT2), and low Ca(2+) electrical fusion with chemical activation (NT3).
- Data on fusion rates, blastocyst cell numbers, recovery rates, and pregnancy rates were collected at multiple gestational stages (Days 6-114).
- Statistical analysis was performed to compare the outcomes between the different protocols.
Main Results:
- Protocols NT1 and NT2 showed significantly higher fusion rates than NT3.
- The NT2 protocol achieved a 100% pregnancy rate, which was significantly higher than NT1.
- All protocols resulted in live, pCAGG-EGFP positive piglets, with no significant differences in recovery rates.
Conclusions:
- The fusion/activation protocol involving electrical fusion followed by MG132 treatment (NT2) demonstrated the highest overall pregnancy rates in porcine SCNT.
- MG132 may enhance SCNT efficiency by stimulating proteasomes within the donor cell nucleus post-transfer.
- This study provides valuable insights into optimizing SCNT techniques for efficient production of genetically engineered pigs.
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