Recent advances in the development of genome editing technologies based on programmable nucleases have substantially improved our ability to make precise changes in the genomes of eukaryotic cells. Genome editing is already broadening our ability to elucidate the contribution of genetics to disease by facilitating the creation of more accurate cellular and animal models of pathological processes. A particularly tantalizing application of programmable nucleases is the potential to directly correct genetic mutations in affected tissues and cells to treat diseases that are refractory to traditional therapies. Here we discuss current progress toward developing programmable nuclease–based therapies as well as future prospects and challenges.

Therapeutic genome editing: prospects and challenges

The ability to manipulate the structure and function of promising nanosystems via energy input and external stimuli is emerging as an attractive paradigm for developing reconfigurable and programmable nanomaterials and multifunctional devices. Light stimulus manifestly represents a preferred external physical and chemical tool for in situ remote command of the functional attributes of nanomaterials and nanosystems due to its unique advantages of high spatial and temporal resolution and digital controllability. Photochromic moieties are known to undergo reversible photochemical transformations between different states with distinct properties, which have been extensively introduced into various functional nanosystems such as nanomachines, nanoparticles, nanoelectronics, supramolecular nanoassemblies, and biological nanosystems. The integration of photochromism into these nanosystems has endowed the resultant nanostructures or advanced materials with intriguing photoresponsive behaviors and more sophisticated functions. In this Review, we provide an account of the recent advancements in reversible photocontrol of the structures and functions of photochromic nanosystems and their applications. The important design concepts of such truly advanced materials are discussed, their fabrication methods are emphasized, and their applications are highlighted. The Review is concluded by briefly outlining the challenges that need to be addressed and the opportunities that can be tapped into. We hope that the review of the flourishing and vibrant topic with myriad possibilities would shine light on exploring the future nanoworld by encouraging and opening the windows to meaningful multidisciplinary cooperation of engineers from different backgrounds and scientists from the fields such as chemistry, physics, engineering, biology, nanotechnology and materials science.

Photochromism into nanosystems: towards lighting up the future nanoworld.

Photoactivatable (alternatively, photoremovable, photoreleasable, or photocleavable) protecting groups (PPGs), also known as caged or photocaged compounds, are used to enable non-invasive spatiotemporal photochemical control over the release of species of interest. Recent years have seen the development of PPGs activatable by biologically and chemically benign visible and near-infrared (NIR) light. These long-wavelength-absorbing moieties expand the applicability of this powerful method and its accessibility to non-specialist users. This review comprehensively covers organic and transition metal-containing photoactivatable compounds (complexes) that absorb in the visible- and NIR-range to release various leaving groups and gasotransmitters (carbon monoxide, nitric oxide, and hydrogen sulfide). The text also covers visible- and NIR-light-induced photosensitized release using molecular sensitizers, quantum dots, and upconversion and second-harmonic nanoparticles, as well as release via photodynamic (photooxygenation by singlet oxygen) and photothermal effects. Release from photoactivatable polymers, micelles, vesicles, and photoswitches, along with the related emerging field of photopharmacology, is discussed at the end of the review.

Visible-to-NIR-Light Activated Release: From Small Molecules to Nanomaterials.

Therapeutic peptides: current applications and future directions

PROTACs: An Emerging Therapeutic Modality in Precision Medicine.

Biological processes are naturally regulated with high spatial and temporal control, as is perhaps most evident in metazoan embryogenesis Chemical tools have been extensively utilized in cell and developmental biology to investigate cellular processes, and conditional control methods have expanded applications of these technologies toward resolving complex biological questions Light represents an excellent external trigger since it can be controlled with very high spatial and temporal precision To this end, several optically regulated tools have been developed and applied to living systems In this review we discuss recent developments of optochemical tools, including small molecules, peptides, proteins, and nucleic acids that can be irreversibly or reversibly controlled through light irradiation, with a focus on applications in cells and animals

Optochemical Control of Biological Processes in Cells and Animals.

Recent advances in the optical control of protein function through genetic code expansion.

Optochemische Steuerung biologischer Vorgänge in Zellen und Tieren

Rapid, easy-to-use, and portable devices that can provide a read-out without the need for expensive equipment represent the future of sensing technology, with applications in areas like environmental, food, chemical, and biological safety. Enzymes immobilized on a surface function as micropumps in the presence of species (e.g., substrate, cofactor, or biomarker) that trigger the enzymatic reaction. The flow speed in these devices increases with increasing reaction rate. This allows the detection of substances that inhibit the enzymatic reaction. Using this principle, sensors for toxic substances, like mercury, cadmium, cyanide, and azide, were designed using urease and catalase-powered pumps, respectively, with limits of detection well below the concentrations permitted by the Environmental Protection Agency. The study was also extended to other inhibitors for these enzymes. The sensing range of fluid flow-based inhibitor assays depends on the type of inhibition, the enzyme concentration on the sensing platform, and, for competitive inhibition, the concentration of substrate used.

Enzyme Micropump‐Based Inhibitor Assays

Protein phosphatases are involved in embryonic development, metabolic homeostasis, stress response, cell cycle transitions, and many other essential biological mechanisms. Unlike kinases, protein phosphatases remain understudied and less characterized. Traditional genetic and biochemical methods have contributed significantly to our understanding; however, these methodologies lack precise and acute spatiotemporal control. Here, we report the development of a light-activated protein phosphatase, the dual specificity phosphatase 6 (DUSP6 or MKP3). Through genetic code expansion, MKP3 is placed under optical control via two different approaches: (i) incorporation of a caged cysteine into the active site for controlling catalytic activity and (ii) incorporation of a caged lysine into the kinase interaction motif for controlling the protein-protein interaction between the phosphatase and its substrate. Both strategies are expected to be applicable to the engineering of a wide range of light-activated phosphatases. Applying the optogenetically controlled MKP3 in conjunction with live cell reporters, we discover that ERK nuclear translocation is regulated in a graded manner in response to increasing MKP3 activity. Protein phosphatases play an essential role in signal transduction, but are understudied due to the difficulties in detecting phosphate removal and the lack of good inhibitors. Here the authors develop a light-activated protein phosphatase using photocaged, unnatural amino acids and use it to study ERK nuclear translocation.

/pdf/optical-control-of-protein-phosphatase-function-407d6suqou.pdf

Taylor M. Courtney

Papers

Optochemical Control of Biological Processes in Cells and Animals.

Recent advances in the optical control of protein function through genetic code expansion.

Optochemische Steuerung biologischer Vorgänge in Zellen und Tieren

Enzyme Micropump‐Based Inhibitor Assays

Optical control of protein phosphatase function