Authors

Maximilian Hörner,1,2,* Carolina Jerez-Longres,1,2,3 Anna Hudek,1,2 Sebastian Hook,1,† O. Sascha Yousefi,1,2,4 Wolfgang W. A. Schamel,1,2,4 Cindy Hörner,5 Matias D. Zurbriggen,6 Haifeng Ye,7 Hanna J. Wagner,1,2,8 and Wilfried Weber1,2,3,*

Affiliations

1Faculty of Biology, University of Freiburg, Freiburg, Germany.
2Signalling Research Centres BIOSS and CIBSS, University of Freiburg, Freiburg, Germany.
3Spemann Graduate School of Biology and Medicine (SGBM), University of Freiburg, Freiburg, Germany.
4Center of Chronic Immunodeficiency CCI, University Clinics and Medical Faculty, Freiburg, Germany.
5Division of Veterinary Medicine, Paul-Ehrlich-Institut, Langen, Germany.
6Institute of Synthetic Biology and CEPLAS, Heinrich Heine University Düsseldorf, Düsseldorf, Germany.
7Synthetic Biology and Biomedical Engineering Laboratory, Biomedical Synthetic Biology Research Center, Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, China.
8Department of Biosystems Science and Engineering, ETH Zurich, Basel, Switzerland.

Topic

Molecular Biology

Abstract

Methodologies for the controlled delivery of genetic information into target cells are of utmost importance for genetic engineering in both fundamental and applied research. However, available methods for efficient gene transfer into user-selected or even single cells suffer from low throughput, the need for complicated equipment, high invasiveness, or side effects by off-target viral uptake. Here, we engineer an adeno-associated viral (AAV) vector system that transfers genetic information into native target cells upon illumination with cell-compatible red light. This OptoAAV system allows adjustable and spatially resolved gene transfer down to single-cell resolution and is compatible with different cell lines and primary cells. Moreover, the sequential application of multiple OptoAAVs enables spatially resolved transduction with different transgenes. The approach presented is likely extendable to other classes of viral vectors and is expected to foster advances in basic and applied genetic research.


DOI: https://dx.doi.org/10.1126%2Fsciadv.abf0797

Extract

For illumination at the confocal microscope, a pE-4000 LED light source (CoolLED, Andover, UK) with 740 nm was used.

Product Associated Features

The pE-4000 Universal Illumination System offers 16 selectable wavelengths from 365 - 770 nm, making it a highly flexible illuminator covering a wide variety of fluorophores

Product Type

pE-4000

Journal

Sci Adv.

Year of Publication

2021

Country of Publication

China, Germany, Switzerland