title: Exogenous Gene Integration for Microalgal Cell Transformation Using a Nanowire-Incorporated Microdevice.
abstract: 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 polydimethylsiloxane (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 × 10(4)- and 9.66 × 10(4)-fold higher than that of a traditional glass bead beating and electroporation.
authors.affiliation: Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST) , 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Sunwoong
authors.initials: S
authors.lastname: Bae
authors.affiliation: Department of Life Science, Research Institute for Natural Sciences, Hanyang University , Seoul 133-791, Republic of Korea.
authors.forename: Seunghye
authors.initials: S
authors.lastname: Park
authors.affiliation: School of Mechanical, Aerospace and Systems Engineering, Division of Mechanical Engineering, Korea Advanced Institute of Science and Technology (KAIST) , 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Jung
authors.initials: J
authors.lastname: Kim
authors.affiliation: Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST) , 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Jong Seob
authors.initials: JS
authors.lastname: Choi
authors.affiliation: Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST) , 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Kyung Hoon
authors.initials: KH
authors.lastname: Kim
authors.affiliation: School of Mechanical, Aerospace and Systems Engineering, Division of Mechanical Engineering, Korea Advanced Institute of Science and Technology (KAIST) , 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Donguk
authors.initials: D
authors.lastname: Kwon
authors.affiliation: Department of Life Science, Research Institute for Natural Sciences, Hanyang University , Seoul 133-791, Republic of Korea.
authors.forename: EonSeon
authors.initials: E
authors.lastname: Jin
authors.affiliation: School of Mechanical, Aerospace and Systems Engineering, Division of Mechanical Engineering, Korea Advanced Institute of Science and Technology (KAIST) , 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Inkyu
authors.initials: I
authors.lastname: Park
authors.affiliation: Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST) , 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Do Hyun
authors.initials: DH
authors.lastname: Kim
authors.affiliation: Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST) , 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Tae Seok
authors.initials: TS
authors.lastname: Seo
mesh_terms: D001709:Biotechnology / Q000379:methods*; D058086:Microalgae; D044085:Microfluidics / Q000379:methods*; D053770:Nanowires / Q000737:chemistry*; D015034:Zinc Oxide / Q000737:chemistry*
chemical_list: D015034:Zinc Oxide
keywords: ZnO nanowire; biofuel; gene delivery; high throughput; microalgal; microfluidics; transformation