The recently discovered CRISPR-Cas gene editing system and its derivatives have found numerous applications in fundamental biology research and pharmaceutical sciences. The need for precise external control over the gene editing and regulatory events has driven the development of inducible CRISPR-Cas systems. While most of the light-controllable CRISPR-Cas systems are based on protein engineering, we developed an alternative synthetic approach based on modification of crRNA/ tracrRNA duplex (guide RNA or gRNA) with photocaging groups, preventing the gRNA from recognizing its genome target sequence until its deprotection is induced within seconds of illumination. This approach relies on a straightforward solid-phase synthesis of the photocaged gRNAs, with simpler purification and characterization processes in comparison to engineering a lightresponsive protein. We have demonstrated the feasibility of photocaging of gRNAs and light-mediated DNA cleavage upon brief exposure to light in vitro. We have achieved light-mediated spatiotemporally resolved gene editing as well as gene activation in cells, whereas photocaged gRNAs showed virtually no detectable gene editing or activation in the absence of light irradiation. Finally, we have applied this system to spatiotemporally control gene editing in zebrafish embryos in vivo, enabling the use of this strategy for developmental biology and tissue engineering applications.
Electrospinning forms fibers from either an electrically charged polymer solution or polymer melt. Over the past decades, it has become a simple and versatile method for nanofiber production. Hence, it has been explored in many different applications. Commonly used electrospinning assembles fibers from polymer solutions in various solvents, known as solution electrospinning, while melt and near-field electrospinning techniques enhance the versatility of electrospinning. Adaption of additive manufacturing strategy to electrospinning permits precise fiber deposition and predefining pattern construction. This manuscript critically presents the potential of electrospun nanofibers in healthcare applications. Research community drew impetus from the similarity of electrospun nanofibers to the morphology and mechanical properties of fibrous extracellular matrices (ECM) of natural human tissues. Electrospun nanofibrous scaffolds act as ECM analogs for specific tissue cells, stem cells, and tumor cells to realize tissue regeneration, stem cell differentiation, and in vitro tumor model construction. The large surface-to-volume ratio of electrospun nanofibers offers a considerable number of bioactive agents binding sites, which makes it a promising candidate for a number of biomedical applications. The applications of electrospinning in regenerative medicine, tissue engineering, controlled drug delivery, biosensors, and cancer diagnosis are elaborated. Electrospun nanofiber incorporations in medical device coating, in vitro 3D cancer model, and filtration membrane are also discussed.
Background The success of biomedical implants in orthopedic and dental applications is usually limited due to insufficient bone‐implant integration, and implant‐related infections. Biointerfaces are critical in regulating their interactions and the desirable performance of biomaterials in biological environment. Surface engineering has been widely studied to realize better control of the interface interaction to further enhance the desired behavior of biomaterials. Purpose and Scope This review aims to investigate surface coating strategies in hard tissue applications to address insufficient osteointegration and implant‐related infection problems. Summary We first focused on surface coatings to enhance the osteointegration and biocompatibility of implants by emphasizing calcium phosphate‐related, nanoscale TiO2‐related, bioactive tantalum‐based and biomolecules incorporated coatings. Different coating strategies such as plasma spraying, biomimetic deposition, electrochemical anodization and LENS are discussed. We then discussed techniques to construct anti‐adhesive and bactericidal surface while emphasizing multifunctional surface coating techniques that combine potential osteointegration and antibacterial activities. The effects of nanotopography via TiO2 coatings on antibacterial performance are interesting and included. A smart bacteria‐responsive titanium dioxide nanotubes coating is also attractive and elaborated. Conclusion Developing multifunctional surface coatings combining osteogenesis and antimicrobial activity is the current trend. Surface engineering methods are usually combined to obtain hierarchical multiscale surface structures with better biofunctionalization outcomes.
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