Related Experiment Video
Updated: Jun 10, 2026
12:13
Zinc-finger Nuclease Enhanced Gene Targeting in Human Embryonic Stem Cells
Published on: August 23, 2014
Efficient, high-throughput transfection of human embryonic stem cells.
Jennifer C Moore1, Kristin Atze, Percy L Yeung
1Stem Cell Research Center and Department of Cell Biology and Neuroscience, Rutgers, The State University of New Jersey, Room D251, 604 Allison Drive, Piscataway, NJ 08854, USA. moore@biology.rutgers.edu
Stem Cell Research & Therapy
|July 28, 2010
Summary
Electroporation systems efficiently transfect human embryonic stem cells (hESC) with nucleic acids, enabling genetic manipulation. These methods offer a reliable, high-throughput approach for research and clinical applications.
Area of Science:
- Stem Cell Biology
- Molecular Biology
- Genetic Engineering
Background:
- Human embryonic stem cells (hESC) are resistant to conventional genetic manipulation techniques.
- Efficient nucleic acid delivery into hESC is crucial for research and clinical applications.
Purpose of the Study:
- To evaluate two commercial electroporation systems for efficient hESC transfection.
- To assess transfection efficiency, cell viability, and pluripotency marker expression.
Main Methods:
- Comparison of Lonza Nucleofection® 96-well Shuttle® System and Invitrogen Neon™ Transfection System.
- Assessment of transfection efficiency via green fluorescent protein expression (flow cytometry).
- Cell viability analysis using an ATP-catalyzed luciferase reaction.
Main Results:
- Both systems achieved high transfection efficiencies with minimal cell viability loss.
- The Nucleofection® system demonstrated superior reproducibility and scalability for high-throughput applications.
- Successful transient and stable transfection of hESC lines, including siRNA delivery for gene knockdown.
Conclusions:
- Electroporation methods offer a reliable and efficient approach for genetic manipulation of hESC.
- These techniques facilitate high-throughput genetic modification of hESC for diverse applications.
