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Emerging digital assessment of biomarkers by linking health-related data obtained from wearable electronic devices and embedded health and fitness sensors in smartphones is opening up the possibility of creating a continuous remote-monitoring platform for disease management. It is considered that the built-in flashlight of smartphones may be utilized to remotely program genetically engineered designer cells for on-demand delivery of protein-based therapeutics. Here, the authors present smartphone-induced insulin release in β-cell line (iβ-cell) technology for traceless light-triggered rapid insulin secretion, employing the light-activatable receptor melanopsin to induce calcium influx and membrane depolarization upon illumination. This iβ-cell-based system enables repeated, reversible secretion of insulin within 15 min in response to light stimulation, with a high induction fold both in vitro and in vivo. It is shown that programmable percutaneous remote control of implanted microencapsulated iβ-cells with a smartphone's flashlight rapidly reverses hyperglycemia in a mouse model of type-1 diabetes., Small, 17 (35), ISSN:1613-6810, ISSN:1613-6829

The ability to remote control the expression of therapeutic genes in mammalian cells in order to treat disease is a central goal of synthetic biology‐inspired therapeutic strategies. Furthermore, optogenetics, a combination of light and genetic sciences, provides an unprecedented ability to use light for precise control of various cellular activities with high spatiotemporal resolution. Recent work to combine optogenetics and therapeutic synthetic biology has led to the engineering of light‐controllable designer cells, whose behavior can be regulated precisely and noninvasively. This Review focuses mainly on non‐neural optogenetic systems, which are often used in synthetic biology, and their applications in genetic programing of mammalian cells. Here, a brief overview of the optogenetic tool kit that is available to build light‐sensitive mammalian cells is provided. Then, recently developed strategies for the control of designer cells with specific biological functions are summarized. Recent translational applications of optogenetically engineered cells are also highlighted, ranging from in vitro basic research to in vivo light‐controlled gene therapy. Finally, current bottlenecks, possible solutions, and future prospects for optogenetics in synthetic biology are discussed., Advanced Science, 6 (1), ISSN:2198-3844

Multigene delivery systems are emerging as key technologies required in diverse research fields in both academia and industry. Examples include genome editing and synthetic biology, cellular reprogramming and stem cell biology, production of complex proteins in structural biology and many others. Many strategies for assembly and delivery of multigene constructs exist but all of them have their limitations. Genetic constructs are typically introduced into mammalian cells by transient transfection. However, the low efficiency of transient transfection and its limited applicability to a variety of important mammalian cells significantly restrict the utility of this approach. Viral systems infect many cell types but usually don’t have enough capacity for multiple genes. Therefore, highly efficient delivery of multigene plasmid in a variety of mammalian cells is a challenge which needs to be addressed. We developed MultiPrime, a baculovirus-mediated multigene expression system, allowing simultaneous expression of several genes from a single virus for mammalian cells. MultiPrime is a modular, non-cytotoxic, non-integrating system and works with a variety of promoters. MultiPrime viruses efficiently transduced a wide range of cell types, including non-dividing primary neurons and induced-pluripotent stem cells (iPS). We showed that MultiPrime can be used for reprogramming, and for genome editing and engineering by CRISPR/Cas9. Moreover, we implemented dual-host-specific cassettes enabling multiprotein expression like full length human antibodies in insect and mammalian cells using a single reagent. Also, MultiPrime-infected Zebrafish embryos showed expression of all expected genes in vivo. Our experiments establish MultiPrime as a powerful and highly efficient tool, to deliver multiple genes for a wide range of applications in primary and established mammalian cells. The thesis is organized in five chapters. The first chapter provides an introduction to available and common strategies for multigene expression in mammalian cells as well as some of the most important applications of multigene delivery in biological research. The chapters two, three, and four present original research reports covering our established system, an optimized protocol necessary for production and utilization of the system and development of some software for analysis of acquired images by our system. In chapter two, Mansouri et al. (2016) present the main project including establishment of MultiPrime and its diverse applications in cell biology. In chapter three, Mansouri et al. (manuscript in preparation) describe an optimized protocol to generate MultiPrime baculovirus particles that harbor complete gene constructs. In chapter four, Rizk et al. (2014 and 2015) present Squassh, Squassh3C and SquasshAnalyst, three user-friendly software modules that enable segmentation and quantification of subcellular structures of fluorescence microscopy images. In the last Chapter, I discuss and summarize our results and refer to the limitations of our system and I propose possible future directions for genetic manipulation of cells.

Optogenetics uses light as a traceless inducer to remotely control cellular behavior with high safety and spatiotemporal precision, and its implementation for therapeutic synthetic biology enable customizable user-defined remedial outputs to be generated from suitably engineered cells. Here, we focus on non-neural optogenetics, describing the tools and strategies available to engineer light-responsive, therapeutic mammalian designer cells and highlighting recent advances in design and translational applications, including cell and gene therapies. We also discuss current limitations in engineering genetically encoded light-sensitive systems and suggest some possible solutions., Current Opinion in Systems Biology, 28

Wearable smart electronic devices, such as smart watches, are generally equipped with green-light-emitting diodes, which are used for photoplethysmography to monitor a panoply of physical health parameters. Here, we present a traceless, green-light-operated, smart-watch-controlled mammalian gene switch (Glow Control), composed of an engineered membrane-tethered green-light-sensitive cobalamin-binding domain of Thermus thermophilus (TtCBD) CarH protein in combination with a synthetic cytosolic TtCBD-transactivator fusion protein, which manage translocation of TtCBD-transactivator into the nucleus to trigger expression of transgenes upon illumination. We show that Apple-Watch-programmed percutaneous remote control of implanted Glow-controlled engineered human cells can effectively treat experimental type-2 diabetes by producing and releasing human glucagon-like peptide-1 on demand. Directly interfacing wearable smart electronic devices with therapeutic gene expression will advance next-generation personalized therapies by linking biopharmaceutical interventions to the internet of things., Nature Communications, 12 (1), ISSN:2041-1723