Chapter 7
3D Nanochannel Electroporation for Macromolecular
Nucleotide Delivery
Lingqian Chang, Chandani Chitrakar, and Mehdi Nouri
Abstract
Delivery of macromolecular nucleotides into the living cells holds a great promise for the development of
new therapeutics. However, its abilities for adoptive immunotherapy, cell reprogramming, and primary cell
transfection have been long-term hindered by the lack of a system that can locally deliver engineered
therapeutic nucleotides (e.g., plasmids, siRNAs, miRNAs) without causing any side effects. In this chapter,
the performance of a novel 3D nanoelectroporation system (3D NEP) is highlighted in three scenarios—
adoptive immunotherapy, cell reprogramming, and adult mouse primary cardiomyocyte transfection.
Detailed protocols were given to introduce the 3D NEP system assembly, as well as their applications in
(1) natural killer (NK) cells transfection by delivery of chimeric antigen receptor (CAR) plasmids; (2) mouse
embryonic fibroblasts transfection with OSKM factors; and (3) miR-29b molecular beacon (BMs) delivery
into primary cardiomyocytes for interrogating the side effect of miR-29b-assisted treatment.
Key words Nanoelectroporation (NEP), Adoptive immunotherapy, Cell reprogramming, Cardiomyocyte transfection, Macromolecule nucleotide
1 Introduction
Macromolecule nucleotides hold great potential in therapeutic areas
ranging from immunotherapy to cell reprogramming for the restoration of disabled cell function [1]. This possibility is attributed to
their properties of high potency, specific binding, and less toxicity
[1]. Intracellular delivery of macromolecular nucleotides enables
direct interrogation and manipulation of genetic activities in living
cells [2]. However, conventional delivery platforms (e.g., lipofectamine, bulk electroporation, viral transduction) face challenges in
transporting large molecules across the cellular membrane with
high efficiency and cellular safety. In this chapter, a novel nanoscale
system, called 3D nanoelectroporation (3D NEP) is highlighted
(Fig. 1). This system achieves high-throughput cell transfection
while offering controllability and uniformity at single-cell resolution.
The unique performance for macromolecular transfection is
Shulin Li et al. (eds.), Electroporation Protocols: Microorganism, Mammalian System, and Nanodevice,
Methods in Molecular Biology, vol. 2050, https://doi.org/10.1007/978-1-4939-9740-4_7,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
69
3D Nanochannel Electroporation for Macromolecular
Nucleotide Delivery
Lingqian Chang, Chandani Chitrakar, and Mehdi Nouri
Abstract
Delivery of macromolecular nucleotides into the living cells holds a great promise for the development of
new therapeutics. However, its abilities for adoptive immunotherapy, cell reprogramming, and primary cell
transfection have been long-term hindered by the lack of a system that can locally deliver engineered
therapeutic nucleotides (e.g., plasmids, siRNAs, miRNAs) without causing any side effects. In this chapter,
the performance of a novel 3D nanoelectroporation system (3D NEP) is highlighted in three scenarios—
adoptive immunotherapy, cell reprogramming, and adult mouse primary cardiomyocyte transfection.
Detailed protocols were given to introduce the 3D NEP system assembly, as well as their applications in
(1) natural killer (NK) cells transfection by delivery of chimeric antigen receptor (CAR) plasmids; (2) mouse
embryonic fibroblasts transfection with OSKM factors; and (3) miR-29b molecular beacon (BMs) delivery
into primary cardiomyocytes for interrogating the side effect of miR-29b-assisted treatment.
Key words Nanoelectroporation (NEP), Adoptive immunotherapy, Cell reprogramming, Cardiomyocyte transfection, Macromolecule nucleotide
1 Introduction
Macromolecule nucleotides hold great potential in therapeutic areas
ranging from immunotherapy to cell reprogramming for the restoration of disabled cell function [1]. This possibility is attributed to
their properties of high potency, specific binding, and less toxicity
[1]. Intracellular delivery of macromolecular nucleotides enables
direct interrogation and manipulation of genetic activities in living
cells [2]. However, conventional delivery platforms (e.g., lipofectamine, bulk electroporation, viral transduction) face challenges in
transporting large molecules across the cellular membrane with
high efficiency and cellular safety. In this chapter, a novel nanoscale
system, called 3D nanoelectroporation (3D NEP) is highlighted
(Fig. 1). This system achieves high-throughput cell transfection
while offering controllability and uniformity at single-cell resolution.
The unique performance for macromolecular transfection is
Shulin Li et al. (eds.), Electroporation Protocols: Microorganism, Mammalian System, and Nanodevice,
Methods in Molecular Biology, vol. 2050, https://doi.org/10.1007/978-1-4939-9740-4_7,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
69