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Transgene delivery via intracellular electroporetic nanoinjection
Aubrey M Wilson1, Quentin T Aten, Nathan C Toone
1Microbiology and Molecular Biology Department, 775 WIDB, Brigham Young University, Provo, UT, 84602, USA.
Transgenic Research
|March 28, 2013
Summary
A new method called intracellular electroporetic nanoinjection (IEN) effectively delivers transgenes into animal cells. This technique, using electrical forces, achieves results comparable to traditional microinjection for creating transgenic animals.
Area of Science:
- Molecular Biology
- Bioengineering
- Developmental Biology
Background:
- Cytoplasmic delivery methods for transgenes have historically faced challenges, limiting their effectiveness compared to pronuclear microinjection.
- Efficient transgene delivery is crucial for creating genetically modified organisms, particularly for research and therapeutic applications.
Purpose of the Study:
- To develop and validate a novel cytoplasmic delivery technique for transgenes, termed intracellular electroporetic nanoinjection (IEN).
- To demonstrate that IEN can achieve comparable success rates to traditional microinjection in producing transgenic animals.
Main Methods:
- Development of a microelectromechanical system (MEMS) device that utilizes electrostatic charge for transgene manipulation and delivery.
- Application of electrical pulses for localized electroporation of transgenes from the cytoplasm into pronuclei.
- Comparative study analyzing viability, transgene integration, and expression rates between IEN and microinjection.
Main Results:
- IEN successfully delivers transgenes into the cytoplasm and subsequently into pronuclei using electrical forces.
- The MEMS device enables localized electroporation within the cytoplasm, overcoming limitations of standard electroporation.
- IEN demonstrated statistically comparable viability, transgene integration, and expression rates to pronuclear microinjection in producing transgenic pups.
Conclusions:
- Intracellular electroporetic nanoinjection (IEN) offers an effective alternative to pronuclear microinjection for generating transgenic animals.
- This technique bypasses the need for visible pronuclei, offering advantages for embryos with reduced pronuclear visibility.
- IEN has the potential to significantly benefit transgenic research by simplifying and improving delivery efficiency.
