Gene editing technologies have seen significant advancements, particularly with the development of adenine base editing (ABE) and CRISPR-associated transposases (CASTs). ABE has been successfully applied to restore CD3δ in hematopoietic stem and progenitor cells from patients with CD3δ severe combined immunodeficiency, achieving a correction rate of 71.2% ± 7.85% (n = 3) (ref: McAuley doi.org/10.1016/j.cell.2023.02.027/). In contrast, CASTs offer a novel approach to targeted DNA integration without creating double-strand breaks, thus minimizing undesirable byproducts associated with traditional CRISPR-Cas9 methods (ref: Lampe doi.org/10.1038/s41587-023-01748-1/). Additionally, the use of interstrand crosslinking of homologous repair templates has been shown to enhance gene editing efficiency by up to fivefold in various human cell types, highlighting the potential for improved outcomes in gene therapy applications (ref: Ghasemi doi.org/10.1038/s41587-022-01654-y/). Moreover, the exploration of thymidine nucleotide metabolism has revealed its critical role in telomere length maintenance, suggesting new avenues for understanding genetic disorders related to telomere dysfunction (ref: Mannherz doi.org/10.1038/s41588-023-01339-5/). This body of work underscores the diverse methodologies being developed to enhance the precision and efficacy of gene editing technologies.

The application of CRISPR technologies in disease treatment has shown promising results, particularly in genetic disorders and cancer. For instance, the use of adenine base editing has been demonstrated to restore T cell generation in patients with CD3δ severe combined immunodeficiency, achieving a significant mutation correction rate (ref: McAuley doi.org/10.1016/j.cell.2023.02.027/). Additionally, prime editing has been utilized to rescue vision in a mouse model of retinitis pigmentosa, showcasing its potential for treating inherited retinal diseases (ref: Qin doi.org/10.1084/jem.20220776/). Furthermore, the identification of thymidine nucleotide metabolism as a key factor in telomere length control opens new pathways for understanding and potentially treating age-related diseases (ref: Mannherz doi.org/10.1038/s41588-023-01339-5/). Moreover, CRISPR technologies have been employed to investigate cancer vulnerabilities, such as the selective targeting of histone demethylase KDM2A in cancers relying on alternative telomere maintenance mechanisms (ref: Li doi.org/10.1038/s41467-023-37480-2/). This multifaceted approach to utilizing CRISPR in disease contexts highlights its versatility and potential for transformative therapies.

Base editing and prime editing represent cutting-edge innovations in the field of genome editing, offering enhanced precision and reduced off-target effects. Recent studies have highlighted the off-target mutations induced by cytosine base editors in transgenic mice, raising concerns about their long-term safety in vivo (ref: Yan doi.org/10.1038/s41467-023-37508-7/). In contrast, prime editing with genuine Cas9 nickases has been shown to minimize unwanted indels, providing a more refined approach to genomic modifications (ref: Lee doi.org/10.1038/s41467-023-37507-8/). Additionally, the development of a new method for synthesizing multiple gRNA libraries facilitates the functional mapping of mammalian genomic regions, further enhancing the capabilities of CRISPR technologies (ref: Pan doi.org/10.1093/nar/). These advancements underscore the ongoing evolution of base and prime editing techniques, which are increasingly being recognized for their potential to address genetic disorders and improve therapeutic outcomes.

Gene therapy has emerged as a transformative approach for treating genetic disorders, with recent advancements showcasing its potential in various applications. The successful application of adenine base editing to restore CD3δ in patients with severe combined immunodeficiency highlights the efficacy of gene therapy in correcting genetic defects (ref: McAuley doi.org/10.1016/j.cell.2023.02.027/). Furthermore, prime editing has been utilized to rescue vision in a mouse model of retinitis pigmentosa, demonstrating its versatility in addressing inherited retinal diseases (ref: Qin doi.org/10.1084/jem.20220776/). The exploration of thymidine nucleotide metabolism as a determinant of telomere length also presents new insights into the genetic factors influencing age-related diseases (ref: Mannherz doi.org/10.1038/s41588-023-01339-5/). Moreover, the identification of PIK3R1 mutations as a cause of activated PI3Kδ syndrome 2 underscores the importance of understanding genetic pathways in the development of targeted therapies for immune dysregulation (ref: Nguyen doi.org/10.1084/jem.20221020/). Collectively, these studies illustrate the rapid progress in gene therapy and its potential to revolutionize treatment strategies for genetic disorders.

CRISPR screening has become an invaluable tool in functional genomics, enabling the identification of genes involved in various biological processes. Recent advancements include the development of organelle-selective click labeling combined with flow cytometry, which allows for high-throughput CRISPR screening of genes involved in phosphatidylcholine metabolism (ref: Tsuchiya doi.org/10.1016/j.cmet.2023.02.014/). Additionally, massively parallel characterization of CRISPR activator efficacy in human induced pluripotent stem cells has revealed how epigenetic features influence transcriptional responses, providing insights into gene regulation during differentiation (ref: Wu doi.org/10.1016/j.molcel.2023.02.011/). Moreover, the synthesis of multiple gRNA libraries through a controlled template-dependent elongation method enhances the ability to pinpoint functional elements within the genome, further advancing the capabilities of CRISPR technologies (ref: Pan doi.org/10.1093/nar/). These innovations in CRISPR screening methodologies are paving the way for deeper understanding of gene function and regulation in various contexts.

The development of effective delivery systems is crucial for the successful application of gene editing technologies. Recent innovations include the use of a bacterial contractile injection system for programmable protein delivery, which offers a novel mechanism for introducing genetic material into eukaryotic cells (ref: Kreitz doi.org/10.1038/s41586-023-05870-7/). Additionally, the synthesis of a combinatorial library of biodegradable ionizable lipids has led to the creation of inhalable delivery vehicles for mRNA and CRISPR-Cas9 gene editors, demonstrating potential for targeted therapies in lung diseases (ref: Li doi.org/10.1038/s41587-023-01679-x/). Furthermore, the ability to tune gene expression through the engineering of upstream open reading frames represents a significant advancement in controlling phenotypic outcomes in crops, showcasing the versatility of gene editing applications (ref: Xue doi.org/10.1038/s41587-023-01707-w/). These advancements in delivery systems are essential for enhancing the efficacy and specificity of gene editing interventions.

As gene editing technologies advance, ethical and safety considerations have become increasingly important. The potential for off-target mutations, particularly with cytosine base editors, raises significant concerns regarding the long-term safety of these tools in therapeutic applications (ref: Yan doi.org/10.1038/s41467-023-37508-7/). Additionally, the reshaping of chromatin architecture by oncogenic factors such as MYC in prostate cancer highlights the need for careful evaluation of gene editing impacts on genomic stability and cancer progression (ref: Wei doi.org/10.1038/s41467-023-37544-3/). These findings underscore the necessity for rigorous safety assessments and ethical frameworks to guide the responsible application of gene editing technologies in clinical settings, ensuring that potential risks are adequately addressed while maximizing therapeutic benefits.