Related Experiment Video
Updated: Nov 3, 2025
05:53
Trans-inner Cell Mass Injection of Embryonic Stem Cells Leads to Higher Chimerism Rates
Published on: May 29, 2018
10.6K
Improving germline transmission efficiency in chimeric chickens using a multi-stage injection approach
Danial Naseri1, Kianoush Dormiani2, Mehdi Hajian1
1Department of Reproductive Biotechnology, Reproductive Biomedicine Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran.
Plos One
|June 4, 2021
Summary
Researchers developed an efficient method for creating transgenic chickens using lentiviral vectors. Injecting vectors into the embryo bloodstream at specific stages significantly improved germline transmission rates for poultry transgenesis.
Area of Science:
- Animal Biotechnology
- Gene Transfer Technologies
- Avian Genetics
Background:
- Poultry transgenesis is crucial for genetic improvement but hindered by low germline transgene transmission efficiency.
- Existing methods for generating transgenic poultry face challenges in achieving efficient and reliable germline integration.
Purpose of the Study:
- To develop and optimize an efficient germline transgenesis method for chickens using a lentiviral vector system.
- To evaluate novel routes and stages for gene transfer to enhance germline transmission in poultry.
Main Methods:
- Utilized a lentiviral vector system for gene transfer in chicken embryos.
- Administered transgenes via multiple injections into embryos at various developmental stages, including bloodstream and chorioallantoic membrane (CAM) vasculature.
- Investigated different injection routes (blastoderm, CAM vasculature, bloodstream) and timing (Hamburger-Hamilton stages 14-15).
Main Results:
- The embryo bloodstream at Hamburger-Hamilton (HH) stages 14-15 yielded the highest single-injection germline transmission (GT) rate of 7.7%.
- Injection into the CAM vasculature achieved a GT efficiency of 2.7%, significantly higher than the 0.4% from blastoderm injection.
- A double-injection strategy combining bloodstream (HH 14-15) and CAM routes resulted in the most efficient method, achieving a 13.6% GT rate.
Conclusions:
- Multiple injections and novel delivery routes, particularly into the bloodstream at HH stages 14-15, significantly enhance germline transmission in poultry.
- The developed lentiviral vector-based method offers an efficient approach for producing transgenic poultry.
- This optimized method holds promise for advancing virus-mediated gene transfer systems in avian transgenesis.
