Exogenous Gene Integration for Microalgal Cell Transformation Using a Nanowire-Incorporated Microdevice

Superior green algal cells showing high lipid production and rapid growth rate are considered as an alternative for the next generation green energy resources. To achieve the biomass based energy generation, transformed microalgae with superlative properties should be developed through genetic engineering. Contrary to the normal cells, microalgae have rigid cell walls, so that target gene delivery into cells is challengeable. In this study, we report a ZnO nanowire-incorporated microdevice for a high throughput microalgal transformation. The proposed microdevice was equipped with not only a ZnO nanowire in the microchannel for gene delivery into cells but also a pneumatic poIydimethylsiloxane (PDMS) microvalve to modulate the cellular attachment and detachment from the nanowire. As a model, hygromycin B resistance gene cassette (Hyg3) was functionalized on the hydrothermally grown ZnO nanowires through a disulfide bond and released into green algal cells, Chlamydomonas reinhardtii, by reductive cleavage. During Hyg3 gene delivery, a monolithic PDMS membrane was bent down, so that algal cells were pushed down toward ZnO nanowires. The supply of vacuum in the pneumatic line made the PDMS membrane bend up, enabling the gene delivered algal cells to be recovered from the outlet of the microchannel. We successfully confirmed Hyg3 gene integrated in microalgae by amplifying the inserted gene through polymerase chain reaction (PCR) and DNA sequencing. The efficiency of the gene delivery to algal cells using the ZnO nanowire-incorporated microdevice was 6.52 X 10(4)- and 9.66 X 10(4)-fold higher than that of a traditional glass bead beating and electroporation.
Publisher
AMER CHEMICAL SOC
Issue Date
2015-12
Language
ENG
Keywords

ZINC-OXIDE NANOWIRES; CHLAMYDOMONAS-REINHARDTII; WALL REMOVAL; ELECTROPORATION; CYANOBACTERIA; BIOMOLECULES; EXPRESSION; LYSIS

Citation

ACS APPLIED MATERIALS INTERFACES, v.7, no.49, pp.27554 - 27561

ISSN
1944-8244
DOI
10.1021/acsami.5b09964
URI
http://hdl.handle.net/10203/205713
Appears in Collection
ME-Journal Papers(저널논문)CBE-Journal Papers(저널논문)
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