PUBLICATION
An alternative method for delivering exogenous material into developing zebrafish embryos
- Authors
- Kohli, V., Robles, V., Cancela, M.L., Acker, J.P., Waskiewicz, A.J., and Elezzabi, A.Y.
- ID
- ZDB-PUB-070711-38
- Date
- 2007
- Source
- Biotechnology and Bioengineering 98(6): 1230-1241 (Journal)
- Registered Authors
- Waskiewicz, Andrew
- Keywords
- zebrafish embryos, femtosecond laser pulses
- MeSH Terms
-
- Animals
- Antigens, Viral/genetics
- DNA, Viral/chemistry
- DNA, Viral/genetics
- DNA, Viral/pharmacokinetics
- Female
- Fluorescein-5-isothiocyanate/pharmacokinetics*
- Fluorescent Dyes/pharmacokinetics
- Immediate-Early Proteins/genetics
- Intracellular Space/metabolism
- Lasers*
- Male
- Nanotechnology/methods
- Permeability
- Quantum Dots
- Transfection/methods
- Zebrafish/embryology*
- Zebrafish/genetics
- PubMed
- 17615558 Full text @ Biotechnol. Bioeng.
Citation
Kohli, V., Robles, V., Cancela, M.L., Acker, J.P., Waskiewicz, A.J., and Elezzabi, A.Y. (2007) An alternative method for delivering exogenous material into developing zebrafish embryos. Biotechnology and Bioengineering. 98(6):1230-1241.
Abstract
Non-invasive manipulation of multicellular systems is important for medical and biological research. The ability to introduce, remove, or modify molecules in the intracellular environment is pivotal to our understanding of cellular structure and function. Herein, we report on an alternative method for introducing foreign material into developing embryos using the application of femtosecond (fs) laser pulses. When intense fs laser pulses are focused to a submicron spot, transient pores are formed, providing a transport pathway for the delivery of exogenous material into embryonic cells. In this study, zebrafish embryos were used as a model system to demonstrate the non-invasiveness of this applied delivery tool. Utilizing optically induced transient pores chorionated and dechorionated zebrafish embryos were successfully loaded with a fluorescent reporter molecule (fluorescein isothiocyanate), Streptavidin-conjugated quantum dots or DNA (Simian-CMV-EGFP). Pore formation was independent of the targeted location, with both blastomere-yolk interface and blastomere pores competent for delivery. Long-term survival of laser manipulated embryos to pec-fin stage was 89 and 100% for dechorionated and chorionated embryos, respectively. To our knowledge, this is the first report of DNA delivery into zebrafish embryos utilizing fs laser pulses.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping