TY - CHAP
T1 - Nanofountain Probe Electroporation for Monoclonal Cell Line Generation
AU - Espinosa, Horacio D.
AU - Mukherjee, Prithvijit
AU - Patino, Cesar
N1 - Publisher Copyright:
© 2020, Springer Science+Business Media, LLC, part of Springer Nature.
PY - 2020
Y1 - 2020
N2 - In the field of genetic engineering, the modification of genes to produce stable cell lines has a variety of applications ranging from the development of novel therapeutics to patient specific treatments. To successfully generate a cell line, the gene of interest must be delivered into the cell and integrated into the genome. The efficiency of cell line generation systems therefore depends on the efficiency of delivery of genetically modifying molecules such as plasmids and CRISPR/CAS9 complexes. In this work, we describe a localized electroporation-based system to generate stable monoclonal cell lines. By employing the nanofountain probe electroporation (NFP-E) system, single cells in patterned cultures are selectively transfected with plasmids, grown, and harvested to obtain stably expressing cell lines. Methods for microcontact printing, cell culture, electroporation, and harvesting are detailed in this chapter.
AB - In the field of genetic engineering, the modification of genes to produce stable cell lines has a variety of applications ranging from the development of novel therapeutics to patient specific treatments. To successfully generate a cell line, the gene of interest must be delivered into the cell and integrated into the genome. The efficiency of cell line generation systems therefore depends on the efficiency of delivery of genetically modifying molecules such as plasmids and CRISPR/CAS9 complexes. In this work, we describe a localized electroporation-based system to generate stable monoclonal cell lines. By employing the nanofountain probe electroporation (NFP-E) system, single cells in patterned cultures are selectively transfected with plasmids, grown, and harvested to obtain stably expressing cell lines. Methods for microcontact printing, cell culture, electroporation, and harvesting are detailed in this chapter.
KW - Cell line generation
KW - Genetic engineering
KW - Localized electroporation
KW - Monoclonal cell lines
KW - Nanofountain probe electroporation
KW - Plasmid transfection
UR - http://www.scopus.com/inward/record.url?scp=85071756114&partnerID=8YFLogxK
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U2 - 10.1007/978-1-4939-9740-4_6
DO - 10.1007/978-1-4939-9740-4_6
M3 - Chapter
C2 - 31468479
AN - SCOPUS:85071756114
T3 - Methods in Molecular Biology
SP - 59
EP - 68
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -