gene editing

Nanocarrier-Based Approaches for Intracellular Protein Transport in Therapeutic Applications

Intracellular protein delivery is becoming a breakthrough tool in biotechnology and medicine, opening new possibilities for gene editing, enzyme replacement, immunomodulation, and targeted therapies. Traditional techniques like microinjection and electroporation can deliver proteins into the cytosol, but their low throughput, risk of cellular damage, high cost, and limited clinical use make them far from ideal. Advances in nanotechnology have introduced a broad range of nanocarrier platforms such as polymeric and lipid nanoparticles, hybrid lipid–polymer systems, dendrimers, extracellular vesicles, and self-assembling protein nanostructures that help overcome many of these limitations. These carriers enhance protein stability, protect them from enzymatic degradation, improve cellular uptake, and enable controlled or stimulus-responsive release. Key features of nanocarriers—including particle size, surface charge, material type, and ligand functionalization—play central roles in how they move within cells and achieve endosomal escape. Nanocarrier systems have proven efficient delivery of therapeutic enzymes, CRISPR-Cas9 ribonucleoprotein complexes, and fluorescent proteins for imaging. Recent innovations such as biomimetic coatings, fluorinated or heterocyclic polymers, and cell-penetrating peptide modifications have further improved delivery precision and biocompatibility. Despite these advances, challenges like cytotoxicity, immune clearance, manufacturing hurdles, and regulatory complexity continue to shape the field. This chapter describes recent nanocarrier technologies in protein delivery nanocarriers, highlights the design principles that underpin effective intracellular transport, and outlines future directions for developing safe, stable, and clinically meaningful protein-based nanomedicines.

Cas12a cleavage and trimming kinetics reveal mismatches as a tool to steer editing

Gene knockouts by CRISPR-Cas nucleases rely on targeted DNA cleavage and error-prone DNA repair: end-joining pathways can introduce insertions and deletions that assist in disrupting the coding sequence. However, only a fraction of edits achieves this, and an unfavorable array of repair outcomes typically requires switching to another editing technology. Key factors that influence repair are the types and lengths of DNA ends following cleavage. Here, we investigated Cas12a's ability to produce different ends and if they can be used to redistribute editing outcomes. We determined the sites and rates of target cleavage by Cas12a in vitro by combining kinetic modeling with nucleotide-resolution assays. For the first time, we show that trimming – repeated cleavage of an already cut target – occurs about 4x faster than initial cleavage; it also presents alternative DNA end structures for cellular repair. We next introduced specific mismatches to the gRNA. Cas12a maintained fast target cleavage, but changed where the target was cleaved and how quickly it was trimmed, compared to matched gRNA. We exploited the differences in cleavage dynamics between matched and mismatched gRNAs to develop reprogrammed gRNAs, i.e. rpgRNAs. Intentionally-mismatched rpgRNAs retained the high editing efficiency observed with traditional gRNAs. However, they redirected editing between in-frame and out-of-frame outcomes to enhance gene knockout success across genes. Reprogrammed gRNAs offer an efficient way to steer editing toward such preferred outcomes, while retaining the simplicity of gene editing with CRISPR-Cas nucleases.

Microbes and Medicine: Revolutionizing Healthcare with Biotech and Pharma

This chapter outlines the significant contributions that pharmaceutical biotechnology and microbiology have made to medication development and healthcare. Using biological systems and living beings, biotechnology creates novel goods, particularly in the medical field. On the other hand, by examining microorganisms essential to manufacturing procedures, pharmaceutical microbiology guarantees the security and effectiveness of medications. Genetic engineering and other contemporary biotechnological techniques have completely changed how diseases are treated and prevented. The chapter emphasizes aseptic processing and contamination control for product safety while highlighting the use of microbes in the production of biopharmaceuticals and biofuels. It is imperative to continue studying microbial genetics since multidrug resistance illnesses are becoming more common. Biotechnology and pharmaceutical microbiology work together to create a symbiotic connection that might lead to revolutionary improvements in healthcare. Cutting-edge methods for managing disease, including gene editing and personalized therapy, provide revolutionary potential. The chapter highlights the critical role these domains play in modern medicine as well as their promise for innovation in the future.

Advances in Organoids for Drug Development: From Discovery to Delivery

Organoid technology has emerged as a promising platform for drug development by enabling the in vitro reconstruction of human tissue architecture, cellular heterogeneity, and key physiological functions. Compared with conventional two-dimensional cell cultures and animal models, organoids provide improved physiological relevance and translational potential. In this review, we systematically summarize the principles of organoid generation, including cell sources, construction strategies, and model classification, and discuss recent advances in integrating organoids with emerging technologies such as microfluidics, gene editing, artificial intelligence, and three-dimensional bioprinting. We further provide a comprehensive overview of organoid applications across the drug development pipeline, including target identification and drug discovery, pharmacodynamic evaluation, absorption, distribution, metabolism, and excretion (ADME) studies, and multi-organ toxicity assessment. Particular emphasis is placed on their expanding role in evaluating advanced drug delivery systems, including transdermal, pulmonary, and nanomedicine-based intelligent delivery platforms, where organoids enable systematic investigation of delivery–penetration–distribution–efficacy–toxicity processes under physiologically relevant conditions. In addition, current challenges, including model standardization, tissue maturation, vascularization, immune integration, and in vitro–in vivo extrapolation, are critically discussed together with future perspectives. Overall, organoid technology is evolving from an emerging experimental model into an integrated platform for drug discovery and translational research, providing new opportunities for more predictive, efficient, and personalized drug development.

Establishing stable non-diapause egg production in p50ma and f38 strains for genome engineering by diapause hormone receptor knockout in Bombyx mori (Lepidoptera: Bombycidae)

Molecular Scissors With a Search Bar: A Contemporary Synthesis Review of CRISPR and the Editing of the Genome

This article presents a narrative review of CRISPR and the Editing of the Genome in the context of Biology. The literature on this topic has expanded substantially over recent decades, yet it remains fragmented across subfields, methods, and national research traditions. Drawing on an interpretive synthesis of representative contributions, the review reconstructs the historical development of the area, examines the conceptual foundations and definitional disputes that organize its debates, and maps the contemporary landscape of research, including the methodological shift toward data-intensive approaches and the institutional pressures that shape publication practice. Particular attention is given to the role of CRISPR and gene editing as organizing themes, and to the conditions under which findings from different research traditions can be brought into productive comparison. The review identifies three synthetic conclusions: the literature is cumulatively strong but organizationally weak; methodological pluralism is better understood as a resource than as a defect; and the growing practical salience of the topic raises the stakes of its unresolved conceptual questions. An agenda for future work is proposed, emphasizing integrative research designs, transparent synthesis practices, and the protection of definitional and infrastructural work on which cumulative progress depends. The article is intended as both a reference map for newcomers and a provocation for specialists in Biology.

Molecular Scissors With a Search Bar: A Critical Survey Review of CRISPR and the Editing of the Genome

This article presents a narrative review of CRISPR and the Editing of the Genome in the context of Biology. The literature on this topic has expanded substantially over recent decades, yet it remains fragmented across subfields, methods, and national research traditions. Drawing on an interpretive synthesis of representative contributions, the review reconstructs the historical development of the area, examines the conceptual foundations and definitional disputes that organize its debates, and maps the contemporary landscape of research, including the methodological shift toward data-intensive approaches and the institutional pressures that shape publication practice. Particular attention is given to the role of CRISPR and gene editing as organizing themes, and to the conditions under which findings from different research traditions can be brought into productive comparison. The review identifies three synthetic conclusions: the literature is cumulatively strong but organizationally weak; methodological pluralism is better understood as a resource than as a defect; and the growing practical salience of the topic raises the stakes of its unresolved conceptual questions. An agenda for future work is proposed, emphasizing integrative research designs, transparent synthesis practices, and the protection of definitional and infrastructural work on which cumulative progress depends. The article is intended as both a reference map for newcomers and a provocation for specialists in Biology.

Stem Cells As A Promising Therapeutic Strategy For Diabetes Mellitus: A Comprehensive Review

Diabetes mellitus is a chronic metabolic disorder characterized by hyperglycemia resulting from impaired insulin secretion, insulin resistance, or both. Insulin and antidiabetic medications can control blood glucose levels well but cannot help recover dysfunctional pancreatic β-cells or change the pathological processes occurring in the body. Stem cell therapy has emerged as a promising regenerative approach that aims to restore pancreatic function and improve glycemic control. This review discusses the therapeutic potential of embryonic stem cells, adult stem cells, mesenchymal stem cells (MSCs), and induced pluripotent stem cells (iPSCs) in diabetes mellitus. Stem cells may act through differentiation into insulin-producing β-like cells, stimulation of endogenous β-cell regeneration, immunomodulation, anti-inflammatory and anti-apoptotic effects, paracrine signaling, and improvement of peripheral insulin sensitivity. Preclinical studies have demonstrated reductions in blood glucose, enhanced insulin secretion, and pancreatic islet regeneration, while early clinical studies have reported improvements in glycemic control, β-cell function, and reduced insulin requirements. However, challenges including immune rejection, tumorigenicity, ethical concerns, limited long-term evidence, high treatment costs, and lack of standardized protocols remain. Future developments involving gene editing, biomaterials, encapsulation, personalized therapies, and artificial intelligence may improve the safety and efficacy of stem cell-based treatments.

Rice leaf structural adjustment to sustain photosynthetic traits under drought and produce more grain: evidence from field studies of the 3K indica panel

Understanding plant traits that contribute to maintenance of physiological activities under drought is crucial for sustainable rice production. Leaf morphological characters together with UAV-based HTP measurements of canopy temperature (CT), NDVI, leaf water potential ( ψ Leaf ), photosynthetic traits like A, g s E and Ci with drought response index (DRI) were collected from >600 indica genotypes in two field dry seasons. This dataset from the 3 K rice sequenced genomes under well-watered (WW) and managed drought stress (MDS) revealed drought-induced leaf morpho-functional interactive changes related to grain yield under drought. Onset of drought caused ~14-20% reduction in Ci , 35 – 56% reduction in A causing serious loss in yield. Correlations analyses from two years field dry seasons dataset revealed that the combination of initially broader leaves capable of increasing leaf thickness in response to drought, as evidenced by changes in SLA, together with maintenance of ψ Leaf , supporting a higher A net can collectively drive towards a higher DRI. The SLA changes up to even ~50% in one of the best performing genotypes with high DRI values upon severe drought stress. GWAS identified seven QTLs for DRI which include known drought-responsive aquaporin, dehydrin, and heat shock protein genes, which coincide with the GWAS interval identified for CT on chromosome 7. This study identified a tentative pathway linking genomic loci to water balance, leaf development, and photosynthesis-related traits under drought. Our results provide opportunities to select the best-performing rice genotypes, enhance understanding about how to improve grain yield of rice under drought by mechanistic exploration whereas aid drought breeding efforts by selecting optimal haplotype combinations or gene edits .

Groundnut Improvement for Aflatoxin Resistance: Progress and Opportunities

ABSTRACT Groundnut, or peanut, is an industrial oilseed crop that serves the food and feed industries and provides income along the value chain. Pre‐ and post‐harvest aflatoxin contamination, caused by Aspergillus spp., hinders the food and feed value and market opportunities of groundnut products. Developing and deploying aflatoxin‐resistant varieties is the most sustainable and economic approach to control aflatoxin for human and animal well‐being. Variable resistance to Aspergillus infection and disease development has been reported, depending on cultivar susceptibility, crop management practices, and environmental conditions. Hence, understanding the physical, biochemical, and genetic basis of resistance mechanisms to Aspergillus infection is vital for the design and deployment of new varieties. Despite modest global efforts, notably in effective aflatoxin diagnosis and identification of the toxic secondary metabolites, there are limited breeding efforts that have bred and deployed aflatoxin‐resistant varieties. This review aims to present the impacts of groundnut aflatoxin contamination and the progress and opportunities in resistance breeding using current technologies and innovations. The first section presents the production status of groundnut and the extent and conditions of aflatoxin contamination. Aflatoxin control methods and components of resistance are described in the second section, followed by progress and opportunities of resistance breeding with advanced technologies, including omics‐assisted and gene‐editing approaches. Information presented in the review may guide breeding and genetic management of aflatoxin, targeting the development of new varieties with desirable product profiles and durable resistance to control aflatoxin contamination along value chains.

Genetic Improvement of the Silkworm (Bombyx mori): From Conventional Breeding to Modern Biotechnology

The silkworm, Bombyx mori, is one of the most economically important insects owing to its exclusive ability to produce high-quality silk fibres that support the global sericulture industry. Genetic improvement of silkworm has been a primary objective of sericulture research for decades to enhance economically important traits including cocoon yield, shell weight, shell ratio, filament length, disease resistance, stress tolerance, fecundity, and silk quality. Traditional breeding approaches based on selection, hybridization, and heterosis exploitation have significantly contributed to the development of high-yielding silkworm breeds. However, the increasing demand for superior silk production under changing climatic conditions necessitates the integration of advanced molecular and genomic technologies into conventional breeding programmes. Recent developments in molecular genetics, quantitative trait locus (QTL) mapping, marker-assisted selection, genome sequencing, transcriptomics, proteomics, and genome editing have considerably improved the understanding of the genetic architecture underlying economically important traits in Bombyx mori. The availability of the complete silkworm genome and advanced bioinformatics tools has accelerated the identification of candidate genes associated with silk protein synthesis, immunity, environmental adaptation, and reproductive performance. Furthermore, CRISPR/Cas9-mediated genome editing and transgenic technologies have opened new opportunities for precise genetic manipulation to improve silk productivity and resistance against pathogens. Despite these advances, challenges including maintenance of genetic diversity, genotype × environment interactions, ethical concerns regarding genetically modified organisms, and climate change continue to influence silkworm breeding programmes. Future research should emphasize the integration of genomics, artificial intelligence, precision breeding, and multi-omics technologies to develop climate-resilient and high-yielding silkworm breeds. This review discusses the principles, methodologies, recent advances, challenges, and future prospects of genetic improvement in silkworm and highlights their significance for sustainable sericulture and global silk production.

Stem Cells As A Promising Therapeutic Strategy For Diabetes Mellitus: A Comprehensive Review

Diabetes mellitus is a chronic metabolic disorder characterized by hyperglycemia resulting from impaired insulin secretion, insulin resistance, or both. Insulin and antidiabetic medications can control blood glucose levels well but cannot help recover dysfunctional pancreatic β-cells or change the pathological processes occurring in the body. Stem cell therapy has emerged as a promising regenerative approach that aims to restore pancreatic function and improve glycemic control. This review discusses the therapeutic potential of embryonic stem cells, adult stem cells, mesenchymal stem cells (MSCs), and induced pluripotent stem cells (iPSCs) in diabetes mellitus. Stem cells may act through differentiation into insulin-producing β-like cells, stimulation of endogenous β-cell regeneration, immunomodulation, anti-inflammatory and anti-apoptotic effects, paracrine signaling, and improvement of peripheral insulin sensitivity. Preclinical studies have demonstrated reductions in blood glucose, enhanced insulin secretion, and pancreatic islet regeneration, while early clinical studies have reported improvements in glycemic control, β-cell function, and reduced insulin requirements. However, challenges including immune rejection, tumorigenicity, ethical concerns, limited long-term evidence, high treatment costs, and lack of standardized protocols remain. Future developments involving gene editing, biomaterials, encapsulation, personalized therapies, and artificial intelligence may improve the safety and efficacy of stem cell-based treatments.