Recent advances of CRISPR-based gene editing technologies and delivery strategies.

Nanoengineering Systems for Gene Therapy: Mechanisms, Modalities, and Future Directions

Nanotechnology has become an important platform in the fields of gene therapy and genome editing, providing delivery strategies that address persistent therapeutic challenges by improving the precision, efficiency, and safety of genetic modifications. This review highlights the central role of nanomaterials in overcoming persistent barriers to genetic interventions, including inefficient delivery, instability of genetic cargo, and off-target effects. Specifically, we emphasize the combined use of nanomaterials with clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) systems, which can improve editing specificity and therapeutic efficacy. Beyond the classical CRISPR/Cas9 platform, this review also discusses next-generation modalities such as base editors, Cas13, prime editing, and the recently described Tandem Interspaced Guide RNA and TIGR-associated protein (TIGR-Tas) system, while considering their therapeutic potential and distinct delivery challenges. By using nanomaterials, the stability and intracellular delivery of genome-editing systems are improved, enabling more effective treatments for genetic disorders and acquired diseases such as cancer and infectious diseases. In addition, nanocarriers provide controlled release, protection from degradation, and better biocompatibility, thereby improving the safety and reliability of gene-editing therapies. Despite these advances, important translational challenges remain, including immunotoxicity, large-scale manufacturing, and regulatory integration. Overall, the continued convergence of nanotechnology and genome engineering may support the development of personalized medicine strategies that adapt genetic engineering tools for patient-specific applications.

In vivo delivery strategies for therapeutic CRISPR genome editing

A comprehensive review of delivery modalities of CRISPR systems solely in vivo that underpin their therapeutic translation and outlines the remaining barriers to durable, tissue-selective, and broadly deployable CRISPR therapeutics is provided.

Nanoparticle-Mediated CRISPR-Cas9 Delivery: Current Strategies, Pharmacological Challenges and Therapeutic Applications in Genetic Disorders

The CRISPR-Cas9 genome-editing system has emerged as a promising technology for the precise correction of disease-causing mutations and has shown significant potential in treating various genetic disorders. However, the successful clinical application of CRISPR-Cas9 largely depends on the development of safe and efficient delivery systems. Among the available approaches, nanoparticle-mediated delivery has gained considerable attention for its low immunogenicity, high biocompatibility, improved targeting, and capacity to deliver diverse CRISPR cargoes. This review summarises the current strategies of nanoparticle-mediated CRISPR-Cas9 delivery, including lipid, polymeric, hybrid, inorganic, and biomimetic nanoparticles. It also discusses major pharmacological challenges such as off-target effects, biodistribution, endosomal escape, immunogenicity, toxicity, and regulatory considerations. Furthermore, recent therapeutic applications in genetic disorders, including sickle cell disease, β-thalassemia, Duchenne muscular dystrophy, cystic fibrosis, hereditary transthyretin amyloidosis, and cancer, are highlighted. Finally, future perspectives on advanced nanocarrier design and clinical translation are discussed. Nanoparticle-mediated CRISPR-Cas9 delivery holds great promise for advancing precision medicine and developing effective therapies for genetic disorders.

CRISPR-Cas9 Technology in Genetic Disorders: Genome Editing Approaches and Therapeutic Applications – A Review

The potential application of CRISPR technology for the possible management of geneticbased conditions, including sickle-cell anemia, β-thalassemia, cystic fibrosis, and Duchenne muscular dystrophy is described.

Highly efficient CRISPR editing enabled by magnetic nanoparticle delivery

Efficient delivery of CRISPR components remains a major determinant of genome editing outcomes. In this study, we compared conventional lipofection with magnetic nanoparticle-assisted gene delivery (magnetofection) for CRISPR-mediated genome editing efficiency using SpCas9 and AsCas12a systems. Based on the average values obtained from multiple independent targets, lipofection resulted in relatively low indel efficiencies, with mean values of average 8.1%–12.47%. In contrast, magnetofection markedly enhanced genome editing outcomes, yielding average indel efficiencies of average 42.29%–45.04%, representing a substantial increase (3.39- and 5.56-fold, respectively) compared with lipofection. This enhancement was consistently observed across both SpCas9-and AsCas12a-mediated editing, indicating that the improved efficiency conferred by magnetic nanoparticle delivery is independent of the nuclease platform. Furthermore, the increased performance of magnetofection was reproducible across multiple genomic loci and cell lines and was also effective under RNP-based delivery conditions, demonstrating its robustness and reliability. In addition to indel-based genome disruption, magnetofection also significantly improved prime editing efficiency (13.95% on average) compared to lipofection (3.81% on average). Overall, our results demonstrate that magnetic nanoparticle-mediated delivery enables highly efficient and reproducible CRISPR genome editing, substantially outperforming conventional lipofection for both indel formation and prime editing. Magnetofection therefore represents a powerful and broadly applicable delivery strategy for next-generation genome editing applications.

Therapeutic CRISPR/Cas9 delivery approaches: strengths, limitations, and a roadmap for clinical translation