Bansal, Tushar and Lenhart, Justin and Kim, Taesung and Duan, Cunming and Maharbiz, Michel M. (2009) Patterned delivery and expression of gene constructs into zebrafish embryos using microfabricated interfaces. Biomedical Microdevices, 11 (3). pp. 633-641.
We demonstrate a method which uses simple microfabrication and microfluidics to produce custom, shaped electroporators for the patterned delivery of foreign molecules into developing embryos. We show how these electroporators can be used to âdrawâ two-dimensional patterns of tracer molecules, DNA and mRNA into the yolk and cells of zebrafish embryos (Danio rerio) at different stages of development. We demonstrate the successful delivery of patterns of Trypan Blue (normal dye), Texas Red (fluorescent dye), GFP-expressing DNA plasmids and GFP expressing mRNA constructs into both chorionated and dechorionated embryos. Both DNA and mRNA were expressed in the desired patterns subsequent to delivery. Square pulses of 10â20Â V (0.20â0.40Â kV/cm), 50â100Â ms width were sufficient to create transient pores and introduce compounds from the late blastula period (3Â hpf) to early pharyngula period (24Â hpf) embryos. Using 24Â hpf dechorionated embryos, we achieved a high survival of 91.3% and 89%, and a delivery efficiency of 38% and 50% for GFP-DNA and GFP-mRNA respectively. Lastly, we demonstrate the simultaneous delivery of different compounds into the developing embryo.
|Subjects:||Material Science > Nanofabrication processes and tools|
Biomedical Science > Nanobiotechnology
Material Science > Nanochemistry
|Deposited On:||25 Aug 2010 11:47|
|Last Modified:||25 Aug 2010 11:47|
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