Toolkit Items

Focused ultrasound (FUS) is a noninvasive physical delivery and control modality that penetrates deep biological tissues and induces confined mild hyperthermia to activate heat-sensitive genetic modules. In the cited 2023 study, FUS was coupled to heat-sensitive CRISPR, CRISPRa, and CRISPRi systems to enable remote spatiotemporal regulation of genome and epigenome function in live cells and animals.

Adeno-associated virus (AAV) is a viral delivery harness used to package and express CRISPR genome-editing components in vivo. In the cited literature, AAV supports single-vector delivery when smaller Cas9 orthologues and their chimeric guide RNAs fit within AAV packaging constraints, enabling robust in vivo genome editing.

Subsequently, we delve into cutting-edge applications of nanoparticles to enhance immune protection, including mosaic and cocktail nanoparticle vaccines, surface-modified targeting strategies, and the integration of mRNA technology with virus-like particles (VLPs).

This review examines recent advancements in nanoparticle( s) (NPs) delivery systems, with a focus on ... lipid nanoparticles (LNPs)... We discussed various NP platforms and their applications, such as ... dry powder formulations of mRNA-loaded LNPs for pulmonary delivery, and LNP-mediated siRNA delivery for respiratory infections.

Deep brain stimulation (DBS) is an established neuromodulation method used as an add-on treatment for severe Parkinson's disease and other chronic neurological conditions. In the cited review, DBS is presented primarily as the clinical benchmark for comparison with optogenetic neuromodulation.

Various CRISPR delivery systems, including viral vectors, nanocarriers, and extracellular vesicles, play crucial roles in the effective access of this tool to neural cells.

non-viral systems such as lipid nanoparticles and engineered exosomes offer lower toxicity and modularity but face targeting limitations

Exosomes possess antigens and immunostimulatory molecules and can serve as cell-free vaccines to induce antitumor immunity. In addition, given their stability, low immunogenicity, and targeting ability, exosomes represent ideal drug delivery systems in tumor immunotherapy.

Emerging delivery vehicles (AAVs, LNPs, lentivirus, virus-like particles) and their translational implications are discussed.

Lipid-polymer hybrid nanoparticles (LPHNPs) are the next-generation nanocarriers that integrate the mechanical strength and sustained-release capacity of polymeric cores with the biocompatibility and high drug-loading efficiency of lipid shells.

To expand the potential use of contrast-enhanced ultrasound beyond intravascular applications, sub-micron agents, often called nanobubbles or ultra-fine bubbles, have recently emerged as a promising tool.

Polyamidoamine (PAMAM) carriers spanning generations G0-G7 were characterized as polymeric nucleic acid delivery formulations for light-directed delivery in vitro using photochemical internalization (PCI). In an OHS EGFP osteosarcoma model, PAMAM G3-G7 supported efficient light-directed siRNA silencing, whereas EGFP mRNA up-regulation was not detected after EGFP mRNA/PAMAM transfection with or without PCI.

Upconversion nanoparticles (UCNPs) are a light-conversion delivery harness that couples near-infrared (NIR) illumination to modules that normally require shorter-wavelength activation. In the cited studies, UCNPs were paired with a UV-responsive triangular DNA nano sucker and with Opto-CRAC to enable NIR-triggered nucleic acid amplification and NIR control of Ca2+-dependent signaling.

The Light Plate Apparatus (LPA) is an open-source light delivery device engineered for 24-well plates that provides two independent optical inputs to each well. It delivers wavelengths spanning 310 to 1550 nm with intensity control over three orders of magnitude and millisecond temporal resolution, and it has been used for spectral and dynamical calibration experiments in optogenetics and photobiology.

increasing clinical experience with in-vivo editing - particularly using lipid nanoparticle (LNP) and adeno-associated virus (AAV)-based platforms - that has also revealed important safety considerations

LITOS is a LED Illumination Tool for Optogenetic Stimulation built from an assembled printed circuit board that controls a commercially available 32×64 LED matrix. It serves as a programmable light-delivery harness for optogenetic stimulation and is reported to support flexible temporal illumination schemes across multiple culture vessel formats.

Ce-doped Gd3(Al,Ga)5O12 (Ce:GAGG) microparticles are injectable yellow-emitting inorganic scintillators used as implanted transducers for X-ray-driven optogenetic control. In the cited study, they converted X-ray irradiation into local light sufficient to activate ChRmine and inhibit via GtACR1, enabling bidirectional modulation of neural activity in mice.

Drug-inducible lentiviral and transposon vectors were used to deliver the PhyB-PIF light-inducible dimerization system together with the synPCB phycocyanobilin synthesis module. In the cited study, doxycycline treatment induced PCB synthesis and enabled PhyB-PIF light-inducible dimerization function.

AAV9-mediated CIB1 transduction is an in vivo gene delivery approach that uses adeno-associated virus serotype 9 to drive CIB1 gain-of-function expression. In hepatocellular carcinoma patient-derived xenografts, this manipulation promoted lenvatinib resistance.

AAV-PA-Cre 3.0 is an adeno-associated viral delivery resource for the photoactivatable Cre recombinase 3.0 system, generated and validated for in vivo mouse applications. It delivers a blue-light-gated Cre/lox recombination system engineered for mammalian expression with reduced background recombination.

Adeno-associated virus (AAV) gene therapy is a promising approach for hemophilia, offering the potential for sustained therapeutic expression of coagulation factors.

Adeno-associated virus (AAV) particles were used as a delivery harness for the BphP1-QPAS1-based TA optogenetic system in neurons. In the cited ChemBioChem study, this application was enabled by the small size of the QPAS1 component.

Adeno-associated virus delivery is a viral gene delivery harness used to deploy the far-red light-induced split-Cre recombinase (FISC) system in vivo. In the cited study, AAV delivery enabled implementation of optogenetically controlled genome engineering in living systems.

Biofunctional nanodot arrays (bNDAs) are nanoscale surface-patterned delivery harnesses designed to spatially control dimerization and clustering of cell-surface receptors. In live cells, they were used to capture extracellularly GFP-tagged Lrp6 and drive assembly of active Wnt signalosomes at the plasma membrane.

This review focuses on blood cell membrane-derived nanocarriers as drug delivery and immune-regenerative platforms.

The CRISPR-Cas9 piggyBac system is a regulatable genome engineering delivery harness reported in human cells. It combines CRISPR-Cas9 with a piggyBac-based system to enable site-specific randomization of the endogenous genome.

We concentrate on three principal bioengineered platforms: (3) drug-eluting and biodegradable stents that convert passive luminal scaffolds into active, long-term drug-releasing devices

Engineered bacteriophages are emerging as a promising class of precision antimicrobials... Advances in synthetic biology and nanotechnology have made it possible to redesign phages with enhanced specificity, expanded functionality, and improved stability, positioning them as versatile tools for microbiota-centered therapies.

Engineered endosymbionts (EES) are engineered intracellular bacteria that escape endosomal destruction, reside in the mammalian cell cytoplasm, and secrete proteins that are transported to the nucleus. In the reported 2021 system, EES were engineered to express mammalian transcription factors and thereby direct host cell gene expression and modulate host cell fate.

Engineered virus-like particles (eVLPs) have emerged as a promising class of delivery systems for genome editing agents.

Exo-nanomaterials, hybrids that fuse EV membranes with synthetic cores, aim to unite EV biocompatibility and trafficking with the loading capacity, modularity and stimulus-responsiveness of engineered nanomaterials.

Fiber optic technologies are described as a delivery harness used to develop implantable devices for generating and recording signals in excitable tissues. In the cited optogenetics context, they support light-based monitoring of biochemical processes and control of cellular activity.

We concentrate on three principal bioengineered platforms: (2) in situ-forming hydrogels that serve as intelligent wound management materials and sustained drug depots

lentiSLiCES is a lentiviral implementation of the self-limiting Cas9 editing system SLiCES. It delivers genome-editing activity to target cells and then promotes self-neutralization by inactivating SpCas9 after editing.

The COVID-19 pandemic marked a turning point in vaccine development, leading to the swift creation of mRNA vaccines delivered via lipid nanoparticles (LNP-mRNA).

the potential of lipid nanoparticle (LNP)-based messenger RNA (mRNA) vaccines to revolutionize HIV prevention

Macrophage transient horizontal gene transfer is a cell-based siRNA delivery harness in which macrophages loaded with siRNA lipoplexes transfer siRNA to cancer cells during in vitro coculture. In the reported system, transferred CIB1-siRNA reduced tumorsphere growth and lowered CIB1 and KI67 mRNA expression in MDA-MB-468 human breast cancer cells.

Material-to-cell synNotch ligand platforms are engineered biomaterial and extracellular matrix systems that present synNotch ligands to mammalian cells. In the reported 2023 implementation, ligands were covalently incorporated into gelatin hydrogels or into cell-generated fibronectin-containing extracellular matrix to activate synthetic Notch receptors and induce programmed transcriptional outputs.

Lipid nanoparticles (LNPs) can overcome these challenges by encapsulating mRNA for protected and efficient delivery to target cells. Importantly, recent advances have demonstrated the potential of mRNA-LNPs to modulate immune cell function with cell-type specificity, enhancing therapeutic precision.

Lipid nanoparticles (LNPs)-validated for potency and safety in COVID-19 mRNA vaccines-offer a versatile, scalable, and immunogenic platform.

The near-infrared activatable protein-conjugated nanoparticle is a nanoparticle-based delivery harness reported for efficient spatially targeted gene editing in brain applications. The available evidence indicates that it combines protein-conjugated nanoparticle delivery with near-infrared activation to localize editing activity.

Recombinant adeno-associated virus (rAAV) is a viral delivery harness used in the cited study to package and deliver an intein-mediated split-Cas9 system for gene therapy. In this context, rAAV enabled cellular delivery of Cas9 fragments that efficiently reconstituted nuclease activity after intein-mediated protein splicing.

The self-sufficient subcutaneous push button-controlled cellular implant is an implantable delivery harness powered by repeated gentle finger pressure on the overlying skin. Finger-pressure actuation deforms an embedded piezoelectric membrane, generates low-voltage electrical energy, and triggers rapid biopharmaceutical release from engineered electro-sensitive human cells.

recent developments in the field of theranostic nanoparticles (TNPs) with dual actions of inhibiting HIF-1a and downstream metabolic targets, while facilitating the imaging and treatment of the tumor

The portable smart blue-light controlled device (PSLC) is an optogenetic delivery harness that provides blue-light input to blue-light-responsive gene modules. In the cited 2024 MED study, it was used to regulate local cytokine expression in the tumor microenvironment.

Species D adenoviruses, such as human adenovirus type 10 (HAdV-D10), are promising candidates due to low seroprevalence in humans... support the advancement of HAdV-D10 as a next-generation platform for gene delivery and vaccine development.

Exosomes derived from chimeric antigen receptor (CAR) T cells preserve antigen specificity and cytotoxic components without the risks of uncontrolled proliferation or cytokine release, offering a safer class of cell free immunotherapies.

The cell membrane biomimetic core-shell system is a membrane-camouflaged delivery harness developed for light-controllable, precise gene editing. In the cited study, it was used as a CRISPR-Cas9 delivery strategy for tumor cell reprogramming and cancer therapy.

The GelMA-Macrophages-LED system is a composite delivery harness composed of a light-crosslinked GelMA hydrogel, gene-modulated macrophages, and a wireless LED device. In the cited study, it was used for in situ light regulation of cardiac inflammation in murine lipopolysaccharide-induced sepsis models, with macrophage photoactivation linked to suppression of inflammatory cytokine production.

Light-emitting diode illumination is a light-delivery harness used with a photoactivatable CRISPR-Cas9 system to provide optical input for temporally and spatially controlled genome editing in mice. In the cited in vivo study, it was applied in the context of embryo implantation and reproductive function.