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Human lactoferrin transgenic rabbits produced efficiently using dimethylsulfoxide-sperm-mediated gene transfer
Lan Li1, Wei Shen, Lingjiang Min
1Institute of Animal Reproduction, Development and Genetic Engineering, Department of Animal Sciences and Technology, Laiyang Agricultural University, Chengyang, Qingdao 266109, China.
Reproduction, Fertility, and Development
|August 26, 2006
Summary
Researchers developed a new method to create transgenic rabbits for producing human lactoferrin (LF) in mammary glands. This technique efficiently generates transgenic offspring and confirms protein expression before birth, aiding bioreactor development.
Area of Science:
- Biotechnology
- Genetics
- Animal Science
Background:
- Transgenic animals are valuable for producing recombinant proteins.
- Assessing mammary gland expression efficiency before offspring birth is challenging.
- Developing reliable methods for evaluating gene expression cassettes is crucial.
Purpose of the Study:
- To establish a simple and efficient method for evaluating mammary gland tissue-expressed gene cassettes.
- To optimize sperm-mediated gene transfer for creating transgenic rabbits.
- To validate the functionality of a gene cassette designed for human lactoferrin expression.
Main Methods:
- Constructed the gene transfer vector pGBC2LF with human lactoferrin (LF) gene under goat beta-casein promoter control.
- Optimized DNA concentration, DMSO concentration, and linear-to-circular DNA ratio for sperm-mediated gene transfer (SMGT).
- Utilized DMSO-sperm-mediated gene transfer (DMSO-SMGT) to produce transgenic rabbits and assessed LF expression in mammary glands.
Main Results:
- Optimized transfection parameters included 30 μg/mL DNA, 3% DMSO, and a 3:1 linear-to-circular DNA ratio.
- Achieved 34.1% and 57.1% transfection efficiency in embryos and offspring, respectively, using DMSO-SMGT.
- 81% of transgenic female rabbits expressed human LF protein in their mammary glands, with levels up to 153 μg/mL.
Conclusions:
- Transgenic rabbits produced via DMSO-SMGT can efficiently express human lactoferrin in mammary glands.
- The developed method allows for early validation of mammary gland expression cassettes.
- This study provides a robust model for developing transgenic bioreactors for therapeutic protein production.
