NDLI: Bio-chip for spatially controlled transfection of nucleic acid payloads into cells in a culture

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Lab on a Chip Vol - 7 Issue - 7

Lab on a Chip Vol - 7 Issue - 6

Lab on a Chip Vol - 7 Issue - 5

Lab on a Chip Vol - 7 Issue - 4

Lab on a Chip Vol - 7 Issue - 3

Lab on a Chip Vol - 7 Issue - 2

Bio-chip for spatially controlled transfection of nucleic acid payloads into cells in a culture

Content Provider	Royal Society of Chemistry (RSC)
Author	Jain, Tilak Muthuswamy, Jit
Copyright Year	2007
Abstract	Transfection of siRNA and plasmid nucleic molecules to animal, microbial and plant cell cultures is a critical process in various research areas, including drug discovery, functional genomics and basic life science research. Till recent times, transfection of these exogenous molecules have been global in nature i.e. targeting all the cells in a culture and lacking capability to spatially confine the transfection to small populations of cells within a single culture. However, in emerging areas like high-throughput screening of large molecule libraries, there is a critical need to transfect multiple different molecules to locally specified regions of a single cell culture and monitor phenotypical changes in these different cell populations. In this study, we present a cell-based biochip that utilizes a microelectrode array to generate localized current density fields that induce electroporation to a targeted group of cells for site-specific transfection of exogenous molecules. More specifically, we optimize the transfection efficiency and viabilities for spatially controlled transfection of Alexa-Fluor-488 conjugated siRNA molecules into NIH3T3 fibroblast cell cultures. Optimal electroporation parameters are established at current density values ranging between 0.05–0.07 µA µm−2 for high transfection efficiencies (>60%) while maintaining viability (>80%) on individual microelectrodes. Additionally, exogenous plasmid molecules are electroporated for site-specific GFP expression and monitored over 48 h in-situ. The microelectrode array technology reported here can therefore be potentially used for targeting specific cells in a culture with spatial precision and transfecting siRNA and plasmids. The microfabrication approach lends itself to significant high-throughput applications in drug-discovery research.
Starting Page	1004
Ending Page	1011
Page Count	8
File Format	HTM / HTML PDF
ISSN	14730197
Volume Number	7
Issue Number	8
Journal	Lab on a Chip
DOI	10.1039/b707479d
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Subscribed
Subject Keyword	Cell culture Small interfering RNA High-throughput screening Electroporation Plasmid Phenotype Transfection Multielectrode array Microfabrication Micrometre Green fluorescent protein Drug discovery Electric current Fibroblast Functional genomics
Content Type	Text
Resource Type	Article
Subject	Chemistry Nanoscience and Nanotechnology Biochemistry Bioengineering Biomedical Engineering

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1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

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