Virus stamping for targeted single-cell infection in vitro and in vivo

Abstract

Genetic engineering by viral infection of single cells is useful to study complex systems such as the brain. However, available methods for infecting single cells have drawbacks that limit their applications. Here we describe 'virus stamping', in which viruses are reversibly bound to a delivery vehicle-a functionalized glass pipette tip or magnetic nanoparticles in a pipette-that is brought into physical contact with the target cell on a surface or in tissue, using mechanical or magnetic forces. Different single cells in the same tissue can be infected with different viruses and an individual cell can be simultaneously infected with different viruses. We use rabies, lenti, herpes simplex, and adeno-associated viruses to drive expression of fluorescent markers or a calcium indicator in target cells in cell culture, mouse retina, human brain organoid, and the brains of live mice. Virus stamping provides a versatile solution for targeted single-cell infection of diverse cell types, both in vitro and in vivo.

Broad gene expression throughout the mouse and marmoset brain after intravenous delivery of engineered AAV capsids

“…The power of directed evolution and AAV engineering to confer novel tropisms and cell-type specificity has broadened potential research applications and enabled new therapeutic approaches in the CNS [43][44][45][46][47][48] . Our results here show that introducing diversity at multiple locations on the capsid surface into native or previously engineered variants can result in useful features, such as increased transduction efficiency and tissue or cell-type specificity in wild-type (WT) mice and marmosets.…”

Genetic Modification of Brain Organoids

“…This can be achieved either through direct visual identification of cells or alternatively by biologically restricting the genetic modification to specific cell types. Visual identification of target cells can be used for single-cell microinjections (Chow et al, 2016;Shull et al, 2019) or approaches such as viral stamping (to be discussed below; Schubert et al, 2018). Biological restriction, on the other hand, makes use of cell type-specific promotors to activate or suppress gene expression (Pasca et al, 2019;Birey et al, 2017).…”

“…Moreover, researchers will in the future not only be limited to the aforementioned approaches using lentiviral vectors, which infect without cell-type specificity, on bulk cell populations or tissue. Rather, selected single cells can be targeted using such viral vectors (Schubert et al, 2018; Figure 2D). This procedure makes use of a technique termed viral stamping, in which viral vectors are brought into direct physical contact with single target cells (Schubert et al, 2018).…”

The first steps in vision: cell types, circuits, and repair

“…We have bound viruses to magnetic nanoparticles and showed that these particles can be remote controlled in the brain using a magnetic field in a way that leads to infection; we named the process "virus stamping" (Schubert et al, 2018). Together with our collaborator Daniel Mueller, we have begun to address this problem.…”

“…Together with our collaborator Daniel Mueller, we have begun to address this problem. We have bound viruses to magnetic nanoparticles and showed that these particles can be remote controlled in the brain using a magnetic field in a way that leads to infection; we named the process "virus stamping" (Schubert et al, 2018).…”