TY - GEN

T1 - Plasmonic-induced molecular transfer and its perspectives in plant Science

AU - Johannsmeier, Sonja

AU - Londenberg, Anke

AU - Zabic, Miroslav

AU - Schiwack, Jana

AU - Boch, Jens

AU - Ripken, Tammo

AU - Heinemann, Dag

N1 - Funding Information: This work is funded by the Federal Ministry of Education and Research, Germany, Grant no. FKZ 031B0542. We gratefully thank Frederik-Matti Bartels for his support during literature and Björn Wünschmann for his major contribution in realizing the GNOME prototype.

PY - 2022

Y1 - 2022

N2 - Molecular transfer across cellular membranes into living cells represents a fundamental technical challenge for the biological sciences. Within the biomedical field, a variety of laser-based transfer methods have been explored and successfully applied. In the most abundant approach, a NIR fs laser is tightly focused on the cellular membrane. However, this approach is limited by the single-cell throughput. Plasmonic nanoparticles have proven as viable mediator to dramatically improve the throughput of laser-based molecular delivery within a process termed gold nanoparticle mediated (GNOME) photoinjection: membrane bound nanoparticles are illuminated by 532 nm, 850 ps laser pulses, leading to a confined nanoheater effect. With careful selection of the process parameters, the effect is localized to tens of nanometers around the nanoparticles and can achieve efficient and gentle transient permeabilization of the cellular membrane using a scanning laser setup. Herein, we investigate GNOME laser transfection in the context of plant cells, which are a promising target for manipulation via genome editing for breeding purposes. For GNOME laser transfection, as well as for other laser-based delivery approaches, the plant's cell wall represents a major barrier for molecules above the size exclusion limit of about 40 to 60 kDa. Permeabilization of the cell wall requires high laser energy, which could raise concerns regarding the viability of the cells. The presented approach therefore includes the formation of protoplasts from climate chamber cultivated Nicotiana benthamiana plants in isotonic solution before incubation with the gold nanoparticles and laser illumination of the sample. We investigate the impact of different process parameters on the viability and delivery efficiency of marker molecules. The presented approach provides the basis for future vector free genome editing of plant cells.

AB - Molecular transfer across cellular membranes into living cells represents a fundamental technical challenge for the biological sciences. Within the biomedical field, a variety of laser-based transfer methods have been explored and successfully applied. In the most abundant approach, a NIR fs laser is tightly focused on the cellular membrane. However, this approach is limited by the single-cell throughput. Plasmonic nanoparticles have proven as viable mediator to dramatically improve the throughput of laser-based molecular delivery within a process termed gold nanoparticle mediated (GNOME) photoinjection: membrane bound nanoparticles are illuminated by 532 nm, 850 ps laser pulses, leading to a confined nanoheater effect. With careful selection of the process parameters, the effect is localized to tens of nanometers around the nanoparticles and can achieve efficient and gentle transient permeabilization of the cellular membrane using a scanning laser setup. Herein, we investigate GNOME laser transfection in the context of plant cells, which are a promising target for manipulation via genome editing for breeding purposes. For GNOME laser transfection, as well as for other laser-based delivery approaches, the plant's cell wall represents a major barrier for molecules above the size exclusion limit of about 40 to 60 kDa. Permeabilization of the cell wall requires high laser energy, which could raise concerns regarding the viability of the cells. The presented approach therefore includes the formation of protoplasts from climate chamber cultivated Nicotiana benthamiana plants in isotonic solution before incubation with the gold nanoparticles and laser illumination of the sample. We investigate the impact of different process parameters on the viability and delivery efficiency of marker molecules. The presented approach provides the basis for future vector free genome editing of plant cells.

KW - GNOME photoinjection

KW - gold nanoparticle

KW - laser transfection

KW - Photoinjection

KW - phytophotonics

KW - plasmon resonance

KW - protoplasts

UR - http://www.scopus.com/inward/record.url?scp=85133645960&partnerID=8YFLogxK

U2 - 10.1117/12.2634131

DO - 10.1117/12.2634131

M3 - Conference contribution

AN - SCOPUS:85133645960

T3 - Proceedings of SPIE - The International Society for Optical Engineering

BT - Nanophotonics IX

A2 - Andrews, David L.

A2 - Bain, Angus J.

A2 - Nunzi, Jean-Michel

PB - SPIE

T2 - Nanophotonics IX 2022

Y2 - 3 April 2022 through 23 May 2022

ER -