Nanofountain Probe Electroporation for Monoclonal Cell Line Generation

Horacio Dante Espinosa*, Prithvijit Mukherjee, Cesar Patino

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc
Pages59-68
Number of pages10
DOIs
StatePublished - Jan 1 2020

Publication series

NameMethods in Molecular Biology
Volume2050
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Fingerprint

Electroporation
Cell Line
Plasmids
Clustered Regularly Interspaced Short Palindromic Repeats
Printing
Genetic Engineering
Genes
Cell Culture Techniques
Genome
Therapeutics

Keywords

  • Cell line generation
  • Genetic engineering
  • Localized electroporation
  • Monoclonal cell lines
  • Nanofountain probe electroporation
  • Plasmid transfection

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Espinosa, H. D., Mukherjee, P., & Patino, C. (2020). Nanofountain Probe Electroporation for Monoclonal Cell Line Generation. In Methods in Molecular Biology (pp. 59-68). (Methods in Molecular Biology; Vol. 2050). Humana Press Inc. https://doi.org/10.1007/978-1-4939-9740-4_6
Espinosa, Horacio Dante ; Mukherjee, Prithvijit ; Patino, Cesar. / Nanofountain Probe Electroporation for Monoclonal Cell Line Generation. Methods in Molecular Biology. Humana Press Inc, 2020. pp. 59-68 (Methods in Molecular Biology).
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Espinosa, HD, Mukherjee, P & Patino, C 2020, Nanofountain Probe Electroporation for Monoclonal Cell Line Generation. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 2050, Humana Press Inc, pp. 59-68. https://doi.org/10.1007/978-1-4939-9740-4_6

Nanofountain Probe Electroporation for Monoclonal Cell Line Generation. / Espinosa, Horacio Dante; Mukherjee, Prithvijit; Patino, Cesar.

Methods in Molecular Biology. Humana Press Inc, 2020. p. 59-68 (Methods in Molecular Biology; Vol. 2050).

Research output: Chapter in Book/Report/Conference proceedingChapter

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Espinosa HD, Mukherjee P, Patino C. Nanofountain Probe Electroporation for Monoclonal Cell Line Generation. In Methods in Molecular Biology. Humana Press Inc. 2020. p. 59-68. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-9740-4_6