Targeted Gene Delivery Research Articles

Spatially-Resolved Organoid Transfection by Porous Silicon-Mediated Optoporation.

Engineering the spatial organisation of organotypic cultures is pivotal for refining tissue models that are useful for gaining deeper insights into complex, non-cell autonomous processes. These advanced models are key to improving the understanding of fundamental biological mechanisms and therapeutic strategies. Controlling gene regulation through spatially-resolved delivery of nucleic acids provides an attractive approach to produce such tissue models. An emerging strategy for spatially-resolved transfection uses photosensitizing nanoparticles coupled with laser pulses to optoporate cells in culture and locally mediate gene delivery. However, localized optoporation in 3D systems remains challenging. Here we propose a solution to this longstanding hurdle, demonstrating that porous silicon nanoparticles are a safe and bioresorbable photosensitising nanomaterial capable of spatially-resolved transfection of mRNA in MCF-7 organoids by near-infrared two-photon optoporation. Functionalization with an azobenzene-lysine photo-switchable moiety enhances the transfection efficiency of the nanoparticles up to 84% in a 2D cell system. Moreover, the nanoparticles enable spatially selective mRNA transfection to MCF-7 spheroids, demonstrating targeted gene delivery in complex 3D cellular environments. The approach for spatially-resolved 3D optoporation offers a way forward for the design of tailored spheroids and organoids by spatially selective nucleic acids delivery.

Aptamer-guided graphene oxide quantum dots for targeted suicide gene therapy in an organoid model of luminal breast cancer

Breast cancer is one of the most common cancers in women. One of the best therapeutic methods against breast cancer is gene therapy, while having an appropriate gene carrier is the biggest challenge of gene therapy. Hence, developing carriers with low cytotoxicity and high gene transfection efficiency, and preferentially with the selective function of gene delivery is a critical demand for this method. In the present study, we introduce a novel targeted carrier to deliver the inducible caspase-9 suicide gene (pLVSIN-iC9) into breast cancer cells. The carrier is composed of graphene oxide quantum dots decorated with polyethyleneimine, and S2.2; an aptamer with high affinity to MUC1 (GOQD-PEI/S2.2). Due to the overexpression of MUC1 in breast cancer cells, the designed GOQD-PEI/S2.2/pLVSIN-iC9 can selectively target cancer cells. Moreover, to better mimic solid tumor conditions, and to evaluate the selective effect of the GOQD-PEI/S2.2/pLVSIN-iC9, an organoid model derived from human dermal fibroblasts (HDF) and MCF-7 cells (coculture organoid) was generated and characterized. The results demonstrate that the coculture organoid model adapts the tissue structure of luminal breast cancer, as well. Therefore, the organoids were subjected to treatment with targeted gene therapy using GOQD-PEI/S2.2/pLVSIN-iC9. Our evidence supports the targeted killing effect of iC9 on the breast cancer cells of the organoids and suggests the good potential of the newly introduced carriers in targeted gene delivery.

Folic-Acid-Terminated Black Phosphorus Nanosheets for Targeted Gene Delivery and Photothermal Therapy against Hepatocellular Carcinoma

Folic-Acid-Terminated Black Phosphorus Nanosheets for Targeted Gene Delivery and Photothermal Therapy against Hepatocellular Carcinoma

Therapeutic Efficacy of Nanocarrier‐Mediated Inhalable Gene Transfection for Lung Cancer

AbstractLung cancer is now the leading cause of cancer‐related mortality, eclipsing all other cancer forms, and of all lung malignancies, non‐small cell lung cancer (NSCLC) makes up around 85%. Recently, the use of vectors in gene therapy besides chemotherapy to treat malignancies brought on by gene mutations has gained prominence. Therapeutic molecule inhalation is a direct approach to lung‐targeted medication delivery with low nonspecific toxicity and limited drug exposure. Treatment for lung cancer with chemotherapy and immunotherapy can be aided by inhalable nanomedicine through advanced mechanisms. Viral and nonviral vectors, such as lipid‐based nanoparticles, polymeric nanoparticles, and inorganic nanoparticles, have all drawn a lot of interest for their ability to increase effects and decrease side effects. Nanocarriers have a significant impact on targeted gene delivery, bioavailability, stability, and residence in target areas. The inhaled pulmonary gene delivery approach, when combined with nanomedicine, will offer a noninvasive and successful way to treat lung cancer by taking use of the physiological properties of the lung. The authors have majorly used data from PubMed and Google Scholar to obtain the relevant information required for the article. In general, this review concentrates on the usage of various inhalable nanocarriers, which might serve as an inspiration for the creation and deployment of more potent genetic therapy for the treatment of lung cancer.

Quantum Dots: An Emerging Approach for Lung Cancer Management

Nanotechnology is an empirical approach that promises hope and novel treatment possibilities for cancer. This state-of-the-art technology offers innovative methods for diagnosing and treating a variety of illnesses. Though there have been some positive advances since the discovery of quantum dots (QD) nano-transporters, these advances are still in their development, even though they have been shown to be beneficial to society. QD has shown to be incredibly suitable for bio-imaging, targeted gene delivery, pharmacological research, biosensing, photodynamic therapy, and diagnosis through its usage in natural imaging and photography. The overview’s main goal was to emphasize how important QD is for both cancer detection and therapy. The goal is to provide readers with a basic understanding of QD, including its benefits and applications in lung cancer management. Furthermore, we have discussed the many studies pertaining to cytotoxic analysis in order to demonstrate the safety of QDs. The current study provides a concise summary of the current research, fabrication methods, and applications of QD in lung cancer treatment.

Biomimetic siRNA nanogels for regulating macrophage polarization and promoting osteogenesis

BackgroundBone fracture regeneration poses significant clinical challenges due to complications such as delayed healing, nonunion, and the limitations of current treatments. ObjectiveThis study introduces a novel therapeutic approach utilizing biomimetic nanogels to silence the Ccl4 gene, aiming to promote bone repair by regulating macrophage polarization. MethodsThe nanogels, composed of tannic acid (TA) and small interfering RNA (siRNA), were designed for targeted gene delivery. ResultsIn vitro findings indicate that siRNA-mediated Ccl4 reduction significantly improves M2 macrophage polarization, which, in turn, promotes osteogenic differentiation of bone marrow-derived mesenchymal stem cells. Increased expression of osteogenic markers and enhanced mineral deposition were observed. The nanogels demonstrated optimal particle size, stability, and cellular uptake, and biocompatibility assays confirmed their non-toxicity. ConclusionThis study underscores the potential of targeted siRNA delivery in modulating immune responses to enhance bone regeneration, offering promising treatment options for complex bone healing scenarios.

Adeno-associated virus serotype 9 antibodies in neonates and young children: Seroprevalence and kinetics

Gene therapies like onasemnogene abeparvovec for spinal muscular atrophy (SMA) utilize adeno-associated virus 9 (AAV9) for targeted gene delivery, which requires an AAV9-antibody (AAV9-Ab) immunoglobulin G (IgG) ≤1:50 titer threshold. This retrospective cohort study evaluated age-related AAV9-Ab IgG seroprevalence for patients with SMA (Part 1), and AAV9-Ab IgG kinetics and time to 1:50 titer threshold in newborns with elevated AAV9-Ab IgG titers (≥1:100) (Part 2). A semi-quantitative ELISA assay was used in Part 1 (N=1,323 patients). For patients aged <12 months, 3.9% (n/N=31/795) had elevated AAV9-Ab IgG titers (≥1:100); prevalence declined with age. In Part 2, a new quantitative ELISA (linear mixed effects model) described continuous AAV9-Ab IgG concentrations for patients with initial titers ≥1:100. AAV9-Ab IgG concentrations waned according to first-order kinetics (58 samples; N=18 patients). The model-based estimation of the AAV9-Ab IgG average half-life was 41 (95% CI: 38–44) days. Based on visualization, 200 EU/mL was a reasonable approximate for the 1:50 titer threshold. In conclusion, initially elevated titers ≥1:100 in newborn patients declined with age. The new quantitative ELISA may allow for quantification of time to threshold for AAV9-Ab IgG retesting for onasemnogene abeparvovec treatment, leading to treatment as early as possible for patients with SMA.

Prospects of Natural Polymers based Nano-drug Delivery Systems in the Treatment of Pulmonary Disorders

Numerous lung conditions, including lung cancer, influenza, acute respiratory distress syndrome, chronic obstructive pulmonary disease [COPD], asthma, and pneumonia, present a great threat to people all over the world. A range of pharmaceutical drugs, peptides, antibodies, and genetic therapies have all been used to treat chronic lung illnesses. Unfortunately, the majority of chronic lung disorders cannot be fully cured by medication alone. At the moment, managing the symptoms is the only asthma treatment. This article provides a brief overview of the state-of-the-art understanding of the function of natural polymeric materials and emphasises recent developments in innovative drug delivery systems that may help treat a variety of lung diseases. Furthermore, the paper also discusses the latest application of natural polymeric materials for targeting gene delivery through different approaches.

A nanobody interaction with SARS-COV-2 Spike allows the versatile targeting of lentivirus vectors.

While investigating methods to target gene delivery vectors to specific cell types, we examined the potential of using a nanobody against the SARS-CoV-2 Spike protein receptor-binding domain to direct lentivirus infection of Spike-expressing cells. Using four different approaches, we found that lentiviruses with surface-exposed nanobody domains selectively infect Spike-expressing cells. Targeting is dependent on the fusion function of the Spike protein, and conforms to a model in which nanobody binding to the Spike protein triggers the Spike fusion machinery. The nanobody-Spike interaction also is capable of directing cell-cell fusion and the selective infection of nanobody-expressing cells by Spike-pseudotyped lentivirus vectors. Significantly, cells infected with SARS-CoV-2 are efficiently and selectively infected by lentivirus vectors pseudotyped with a chimeric nanobody protein. Our results suggest that cells infected by any virus that forms syncytia may be targeted for gene delivery by using an appropriate nanobody or virus receptor mimic. Vectors modified in this fashion may prove useful in the delivery of immunomodulators to infected foci to mitigate the effects of viral infections.IMPORTANCEWe have discovered that lentiviruses decorated on their surfaces with a nanobody against the SARS-CoV-2 Spike protein selectively infect Spike-expressing cells. Infection is dependent on the specificity of the nanobody and the fusion function of the Spike protein and conforms to a reverse fusion model, in which nanobody binding to Spike triggers the Spike fusion machinery. The nanobody-Spike interaction also can drive cell-cell fusion and infection of nanobody-expressing cells with viruses carrying the Spike protein. Importantly, cells infected with SARS-CoV-2 are selectively infected with nanobody-decorated lentiviruses. These results suggest that cells infected by any virus that expresses an active receptor-binding fusion protein may be targeted by vectors for delivery of cargoes to mitigate infections.

Targeted Gene Delivery to Muscle Cells In Vitro and In Vivo Using Electrostatically Stabilized DNA—Peptide Complexes

Genetic constructs must be delivered selectively to target tissues and intracellular compartments at the necessary concentrations in order to achieve the maximum therapeutic effect in gene therapy. Development of targeted carriers for non-viral delivery of nucleic acids into cells, including those in muscle, which is one of the most challenging tissues to transfect in vivo, remains a topical issue. We have studied ternary complexes of plasmid DNA and an arginine–histidine-rich peptide-based carrier coated with a glutamate–histidine-rich polymer bearing skeletal muscle targeting peptide (SMTP) for the gene delivery to muscle tissue. The relaxation of the ternary complexes after polyanion treatment was assessed using the ethidium bromide displacement assay. The developed polyplexes were used to transfect C2C12 myoblasts in full-media conditions, followed by analysis of their toxic properties using the Alamar Blue assay and expression analysis of lacZ and GFP reporter genes. After delivering plasmids containing the GFP and lacZ genes into the femoral muscles of mdx mice, which are model of Duchenne muscular dystrophy, GFP fluorescence and β-galactosidase activity were detected. We observed that the modification of ternary polyplexes with 10 mol% of SMTP ligand resulted in a 2.3-fold increase in lacZ gene expression when compared to unmodified control polyplexes in vivo. Thus, we have demonstrated that the developed DNA/carrier complexes and SMTP-modified coating are nontoxic, are stable against polyanion-induced relaxation, and can provide targeted gene delivery to muscle cells and tissues. The results of this study are useful for a range of therapeutic applications, from immunization to amelioration of inherited neuromuscular diseases.

CpG Oligodeoxynucleotide-Coated Chitosan Nanoparticles Enhance Macrophage Proinflammatory Phenotype In Vitro.

This study aimed to investigate the effects of CpG Oligodeoxynucleotide (CpG ODNs)-Coated Chitosan Nanoparticles (CNP) on the phenotype of murine macrophages and their pro-inflammatory cytokine profile in vitro. CNP-CpG ODNs loaded with FITC-scrambled siRNA were prepared using the ionotropic gelation method. Peritoneal macrophages were isolated and exposed to CNP-CpG ODNs. Treated macrophages were assessed for uptake capacity. Flow cytometry was used to evaluate the expression levels of MHC-II, CD40, and CD86 costimulatory molecules in treated macrophages. Furthermore, the secretion levels of proinflammatory cytokines (TNF-α and IL-6) and the release of nitric oxide (NO) were measured in the culture supernatant of treated macrophages using sandwich ELISA and the Griess reaction, respectively. These in vitro studies showed that CNP-CpG ODNs had no cytotoxic effect on macrophages and were efficiently taken up by them. Additionally, CNP-CpG ODNs significantly increased the production of TNF-α, IL-6, and NO in the culture supernatant compared to CNP alone. Moreover, CNP-CpG ODNs enhanced the expression of MHC-II, CD40, and CD86 costimulatory molecules on macrophages. These findings indicate that incorporating CpG ODNs into CNPs promotes macrophage maturation and a proinflammatory phenotype. Therefore, CNP-CpG ODNs may serve as an effective system for targeted gene delivery to macrophages, enhancing immune responses.

Layer-by-layer nanocomposite based on zein and hyaluronic acid for tumor-targeted gene delivery

Gene vectors provide a viable approach to ensuring that therapeutic genes reach the target cells. Polyethyleneimine (PEI), a cationic polymer, was brought into focus on high transfection efficiency as a gene carrier. However, the high cytotoxicity and lack of targeting ability during loading genes remain confusing topics. Herein, Fe3O4 served as the magnetic source, plant extract Zein served as the structural unit loaded with PEI and hyaluronic acid (HA) served as the targeted switch, the Fe3O4/Zein/PEI25k-HA (FeZP25k-HA) was synthesized through an antisolvent approach, and its cytotoxicity, targeting ability and transfection efficiency in HEK 293 T and HeLa cells were further investigated. The FeZP25k-HA exhibited low toxicity at the cellular level, and under external magnetic fields, it revealed a significant enrichment effect, which contributed to enhancing cellular uptake. The FeZP25k (Fe3O4/Zein/PEI25k) modified with HA displayed higher uptake and transfection efficiency in HeLa cells, and excellent tumor targeting ability. This work was expected to offer an innovative strategy to design a safe, efficient magnetic targeted gene delivery system with low toxicity and high transfection efficiency.

Ultrasound-Sensitive Targeted Liposomes as a Gene Delivery System for the Synergistic Treatment of Hepatocellular Carcinoma.

Gene therapy and sonodynamic therapy, as emerging treatment methods, have great potential in cancer treatment. However, there are significant challenges in the in vivo delivery of genes and sonosensitizers during the treatment process, which ultimately affects the therapeutic outcome. In this study, an ultrasound-sensitive targeted liposome nanoparticle system (MLipsiBcl-2) is developed to deliver the sonosensitizers and siRNA for the synergistic treatment of hepatocellular carcinoma. Generation of reactive oxygen species (ROS) by MLipsiBcl-2 can be initiated through ultrasound stimulation, leading to liposome rupture and release of the sonosensitizer and small interfering RNA (siRNA). Furthermore, ROS can disrupt lysosomal membranes, facilitating gene release for downregulating overexpressed antiapoptotic protein levels in cancer cells. Experimental results from in vitro and in vivo studies demonstrated the efficacy of synergistic treatment on hepatocellular carcinoma cells and the high biocompatibility of MLipsiBcl-2 under ultrasound stimulation. The advancement of this ultrasound-sensitive targeted gene delivery system shows potential as a versatile therapeutic platform that is easily operable, presenting a prospect for a synergistic treatment approach across various cancer types.

Biosurfactant Nanomicelles and Peptide Integration: Novel Approaches to Targeted Gene Delivery in Colon Cancer Treatment.

A notable breakthrough in the treatment of colon cancer involves the utilisation of a cutting-edge drug delivery technology known as biosurfactant-derived nanomicelles. These nanomicelles, composed of natural biosurfactant molecules, possess the distinct capability to enclose pharmaceuticals or genetic material, such as DNA, siRNA, or mRNA, within spherical formations. With a size ranging from 10 to 100 nanometers, these nanomicelles exhibit precision targeting capabilities towards colon cancer cells, hence minimising the occurrence of side effects typically associated with treatment. Upon being specifically targeted, the nanomicelles liberate their cargo into cancer cells, resulting in enhanced therapy efficacy. This novel strategy utilises the specific attributes of the tumour microenvironment to administer precise and focused treatment. These nanomicelles improve the absorption by cells and reduce harm to healthy tissues by imitating important nutrients or utilising compounds that specifically target tumours. Furthermore, the incorporation of stimuli-responsive components allows for regulated medication release in reaction to the acidic environment seen in tumours. The review focuses on examining the use of biosurfactants and natural peptides in nanomicellar carriers as ways to fight against colon cancer. Folate-coated nanomicelles incorporating curcumin facilitate precise gene delivery, while the partnership of biosurfactants, such as surfactin from Bacillus subtilis and natural peptides, enables the transportation of particular cyclopeptides into the tumour network. Peptides, similar to bombesin, direct nanomicelles to specific places, while peptides based on curcumin control the release of medicinal substances. While preclinical investigations demonstrate promise, obstacles remain in formulation and regulatory issues. However, biosurfactant-based nanomicelles, particularly folate-coated carriers loaded with curcumin, show tremendous potential in overcoming biological barriers and delivering medicines efficiently to colon cancer cells.

Bioprinting of osteochondral tissues: A perspective on current gaps and future trends.

Osteochondral tissue regeneration has remained a critical challenge in orthopaedic surgery, especially due to complications of arthritic degeneration arising out of mechanical dislocations of joints. The common gold standard of autografting has several limitations in presenting tissue engineering strategies to solve the unmet clinical need. However, due to the complexity of joint anatomy, and tissue heterogeneity at the interface, the conventional tissue engineering strategies have certain limitations. The advent of bioprinting has now provided new opportunities for osteochondral tissue engineering. Bioprinting can uniquely mimic the heterogeneous cellular composition and anisotropic extra-cellular matrix (ECM) organization, while allowing for targeted gene delivery to achieve heterotypic differentiation. In this perspective, we discuss the current advances made towards bioprinting of composite osteochondral tissues and present an account of challenges—in terms of tissue integration, long-term survival, and mechanical strength at the time of implantation—required to be addressed for effective clinical translation of bioprinted tissues. Finally, we highlight some of the future trends related to osteochondral bioprinting with the hope of in-clinical translation.

Construction of intelligent response gene vector based on MOF/Fe3O4/AuNRs for tumor-targeted gene delivery

Metal-organic frameworks (MOFs) have the potential to efficiently carry cargo due to their excellent porosity and high surface area. Nevertheless, conventional MOFs and their derivatives exhibit low efficiency in transporting nucleic acids and other small molecules, as well as having poor colloidal stability. In this study, a ZIF-90 loaded with iron oxide nanoparticles and Au nanorods was prepared, and then surface-functionalized with polyethyleneimine (PEI) to create a multifunctional nanocomposite (AFZP25k) with pH, photothermal, and magnetic responsiveness. AFZP25k can condense plasmid DNA to form AFZP25k/DNA complexes, with a maximum binding efficiency of 92.85 %. DNA release assay showed significant light and pH responsiveness, with over 80 % cumulative release after 6 h of incubation. When an external magnetic field is applied, the cellular uptake efficiency in HeLa cells reached 81.51 %, with low cytotoxicity and specific distribution. In vitro transfection experiments demonstrated a gene transfection efficiency of 44.77 % in HeLa cells. Following near-infrared irradiation, the uptake efficiency and transfection efficiency of AFZP25k in HeLa cells increased by 21.3 % and 13.59 % respectively. The findings indicate the potential of AFZP25k as an efficient and targeted gene delivery vector in cancer gene therapy.

Targeted gene therapy for rare genetic kidney diseases

Chronic kidney disease (CKD) is one of the leading causes of mortality worldwide because of kidney failure and the associated challenges of its treatment including dialysis and kidney transplantation. About one-third of CKD cases are linked to inherited monogenic factors, making them suitable for potential gene therapy interventions. However, the intricate anatomical structure of the kidney poses a challenge, limiting the effectiveness of targeted gene delivery to the renal system. In this review, we explore the progress made in the field of targeted gene therapy approaches and their implications for rare genetic kidney disorders, examining preclinical studies and prospects for clinical application. In vivo gene therapy is most commonly used for kidney-targeted gene delivery and involves administering viral and non-viral vectors through various routes such as systemic, renal vein and renal arterial injections. Small nucleic acids have also been used in preclinical and clinical studies for treating certain kidney disorders. Unexpectedly, hematopoietic stem and progenitor cells have been used as an ex vivo gene therapy vehicle for kidney gene delivery, highlighting their ability to differentiate into macrophages within the kidney, forming tunneling nanotubes that can deliver genetic material and organelles to adjacent kidney cells, even across the basement membrane to target the proximal tubular cells. As gene therapy technologies continue to advance and our understanding of kidney biology deepens, there is hope for patients with genetic kidney disorders to eventually avoid kidney transplantation.

A pseudotyped adenovirus serotype 5 vector with serotype 49 fiber knob is an effective vector for vaccine and gene therapy applications

Adenoviruses (Ad) have demonstrated significant success as replication-deficient (RD) viral vectored vaccines, as well as broad potential across gene therapy and cancer therapy. Ad vectors transduce human cells via direct interactions between the viral fiber knob and cell surface receptors, with secondary cellular integrin interactions. Ad receptor usage is diverse across the extensive phylogeny. Commonly studied human Ad serotype 5 (Ad5), and chimpanzee Ad-derived vector “ChAdOx1” in licensed ChAdOx1 nCoV-19 vaccine, both form primary interactions with the Coxsackie and Adenovirus Receptor (CAR), which is expressed on human epithelial cells and erythrocytes. CAR usage is suboptimal for targeted gene delivery to cells with low/negative CAR expression, including human dendritic cells (DCs) and vascular smooth muscle cells (VSMCs). We evaluated the performance of a RD Ad5 vector pseudotyped with the fiber knob of human Ad serotype 49, termed Ad5/49K vector. Ad5/49K demonstrated superior transduction of murine and human DCs over Ad5, which translated into significantly increased T cell immunogenicity when evaluated in a mouse cancer vaccine model using 5T4 tumour-associated antigen. Additionally, Ad5/49K exhibited enhanced transduction of primary human VSMCs. These data highlight the potential of Ad5/49K vector for both vascular gene therapy applications and as a potent vaccine vector.

Nanoliposomes as nonviral vectors in cancer gene therapy.

Nonviral vectors, such as liposomes, offer potential for targeted gene delivery in cancer therapy. Liposomes, composed of phospholipid vesicles, have demonstrated efficacy as nanocarriers for genetic tools, addressing the limitations of off-targeting and degradation commonly associated with traditional gene therapy approaches. Due to their biocompatibility, stability, and tunable physicochemical properties, they offer potential in overcoming the challenges associated with gene therapy, such as low transfection efficiency and poor stability in biological fluids. Despite these advancements, there remains a gap in understanding the optimal utilization of nanoliposomes for enhanced gene delivery in cancer treatment. This review delves into the present state of nanoliposomes as carriers for genetic tools in cancer therapy, sheds light on their potential to safeguard genetic payloads and facilitate cell internalization alongside the evolution of smart nanocarriers for targeted delivery. The challenges linked to their biocompatibility and the factors that restrict their effectiveness in gene delivery are also discussed along with exploring the potential of nanoliposomes in cancer gene therapy strategies by analyzing recent advancements and offering future directions.

Engineering viral vectors for acoustically targeted gene delivery

Targeted gene delivery to the brain is a critical tool for neuroscience research and has significant potential to treat human disease. However, the site-specific delivery of common gene vectors such as adeno-associated viruses (AAVs) is typically performed via invasive injections, which limit its applicable scope of research and clinical applications. Alternatively, focused ultrasound blood-brain-barrier opening (FUS-BBBO), performed noninvasively, enables the site-specific entry of AAVs into the brain from systemic circulation. However, when used in conjunction with natural AAV serotypes, this approach has limited transduction efficiency and results in substantial undesirable transduction of peripheral organs. Here, we use high throughput in vivo selection to engineer new AAV vectors specifically designed for local neuronal transduction at the site of FUS-BBBO. The resulting vectors substantially enhance ultrasound-targeted gene delivery and neuronal tropism while reducing peripheral transduction, providing a more than ten-fold improvement in targeting specificity in two tested mouse strains. In addition to enhancing the only known approach to noninvasively target gene delivery to specific brain regions, these results establish the ability of AAV vectors to be evolved for specific physical delivery mechanisms.