Magnetically guided virus stamping for the targeted infection of single cells or groups of cells
Abstract
To understand and control complex tissues, the ability to genetically manipulate single cells is required. However, current delivery methods for the genetic engineering of single cells, including viral transduction, suffer from limitations that restrict their application. Here we present a protocol that describes a versatile technique that can be used for the targeted viral infection of single cells or small groups of cells in any tissue that is optically accessible. First, cells of interest are selected using optical microscopy. Second, a micropipette—loaded with magnetic nanoparticles to which viral particles are bound—is brought into proximity of the cell of interest, and a magnetic field is applied to guide the viral nanoparticles into cellular contact, leading to transduction. The protocol, exemplified here by stamping cultured neurons with adeno-associated viruses (AAVs), is completed in a few minutes and allows stable transgene expression within a few days, at success rates that approach 80%. We outline how this strategy is applied to single-cell infection in complex tissues, and is feasible both in organoids and in vivo.
Additional Information
© 2019 Springer Nature Limited. Received 10 October 2018; Accepted 02 July 2019; Published 18 October 2019. Data availability: The data shown in this protocol are available upon reasonable request. We thank P. Buchmann and P. Argast for building our magnet Supplementary Figure 1. The study was supported by an European Union grant (FP7/211800 to D.J.M.), a Canada Research Chair (Tier II to S.T.), Swiss National Science Foundation grants (310030B_160225 to D.J.M. and 3100330B_163457 to B.R.), the National Center of Competence in Research (NCCR) Molecular Systems Engineering (to D.J.M. and B.R.), the European Research Council (669157, RETMUS to B.R.) and a DARPA grant (HR0011-17-C-0038, Cortical Sight to B.R.). Author Contributions: All authors discussed the protocol and contributed to the writing of the protocol. R.S., S.H. and S.T. designed and optimized the experiments described. Figures and videos were created by R.S., S.H. and S.T. Competing interests: R.S., S.T., D.J.M. and B.R. applied for a patent related to the virus-stamping approach. The remaining author declares no competing interests.Attached Files
Supplemental Material - 41596_2019_221_Fig6_ESM.jpg
Supplemental Material - 41596_2019_221_MOESM1_ESM.pdf
Supplemental Material - 41596_2019_221_MOESM2_ESM.pdf
Supplemental Material - 41596_2019_221_MOESM3_ESM.mp4
Supplemental Material - 41596_2019_221_MOESM4_ESM.mov
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Additional details
- Eprint ID
- 99518
- Resolver ID
- CaltechAUTHORS:20191028-161515958
- FP7/211800
- European Union
- Canada Research Chairs Program
- 310030B_160225
- Swiss National Science Foundation (SNSF)
- 3100330B_163457
- Swiss National Science Foundation (SNSF)
- 669157
- European Research Council (ERC)
- HR0011-17-C-0038
- Defense Advanced Research Projects Agency (DARPA)
- Created
-
2019-10-29Created from EPrint's datestamp field
- Updated
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2021-11-16Created from EPrint's last_modified field