An encoded viral micropatch for multiplex cell-based assays through localized gene delivery

Abstract

The increasing number of potential drug targets and compounds has led to the development of high-throughput cell-based assays. Simultaneous processing of multiple targets in the same experiment based on localized target gene expression is a very efficient strategy for this purpose. To address this need, we present an adenoviral vector-immobilized microparticle with two-dimensional (2D) shape-encoding properties that allows localized patch-like gene delivery to monolayer-cultured cells. This format conveniently achieves multiplexed gene delivery compatible with both high-throughput cellular assays and fluorescence high-content imaging instruments. A multiplex G protein-coupled receptor (GPCR) internalization assay was developed to demonstrate the compatibility of this system with high-throughput image-based cellular assays.

One‐Step Generation of a Drug‐Releasing Hydrogel Microarray‐On‐A‐Chip for Large‐Scale Sequential Drug Combination Screening

“…We envision that personalized screening results from sequential combinations will help determine drug schedules for each patient . In addition, our platform for multistep combinatorial screening can potentially be applied in other research fields, such as the differentiation and reprogramming of various cell types, the performance of drug screening using cells transduced with various viral vectors, or the creation of personalized drug scheduling using patient‐derived cell lines. We expect that multistep, large‐scale screening using partipetting will be more accessible to researchers in a wide range of fields, thereby broadening the applications of high‐throughput, sequence‐dependent combinatorial bioassays.…”

Emerging barcode particles for multiplex bioassays

“…Another "shape-encoded viral micropatch" also has been developed for convenient localization of gene delivery in multiplex, high-content cellular assays (Fig. 6c) [131]. The conditional transduction of biotinylated adenoviral vectors allows strict confinement of adenoviral gene delivery under the micropatches, and the micropatch is also suitable for image-based assays after the gene delivery, but not so much for performing molecular analysis, by collecting cells after the assays.…”

Hierarchical shape-by-shape assembly of microparticles for micrometer-scale viral delivery of two different genes

“…Here, we describe the shape-by-shape assembly of multiple virus-laden compartments inside microscale virus-laden particles to form a heterogeneous population of cells using microscale transfection, which builds on our previous report on viral micropatches. 7 By assembling daughter microparticles with different shapes and sizes to a parent particle, which has indentations that match the size and shape of the daughter particles [Figs. 1(a)], we were able to demonstrate two different populations of cells in direct contact on a single particle.…”

“…Powerful approaches, such as mechanical cell contacting methods, 5 patterning different materials for different types of cells, 6 and selective transfection, 7 have been developed to address this issue. The systems that use macroscale systems, however, are not only limited in their flexibility in design but also restricted to millimeter scale, which are far from genuine in vivo systems.…”