How Gene Editing Can Support Sustainable Agriculture

BackgroundThe views of people with genetic conditions are crucial to include in public dialogue around developing gene editing technologies. This qualitative study sought to characterize the attitudes of people with inherited retinal conditions (retinitis pigmentosa [RP] and Leber congenital amaurosis [LCA]) toward gene editing.MethodsIndividuals with RP (N = 9) and LCA (N = 8) participated in semi‐structured qualitative interviews about their experience with and attitudes toward blindness, and their views about gene editing technology for somatic, germline, and enhancement applications.ResultsParticipants saw potential benefits from gene editing in general, but views about its use for retinal conditions varied and were influenced by personal perspectives on blindness. Those who felt more negatively toward blindness, particularly those with later onset blindness, were more supportive of gene editing for retinal conditions. Concerns about both germline and somatic editing included: the importance of informed consent; impacts of gene editing on social attitudes and barriers affecting blind people; and worries about “eliminating” blindness or other traits.ConclusionPeople with RP and LCA have diverse attitudes toward gene editing technology informed by their own lived experience with disability, and many have concerns about how the ways in which it is discussed and implemented might affect them.

CRISPR-Cas9 studies have implicated gene editing of DNA sequences for medical treatment of diseases such as sickle cell anemia, thalassemia, AIDS, blindness, muscular dystrophy, Huntington’s disease, and cystic fibrosis. Moreover, gene editing technology has contributed to the detection of viruses responsible for the dissemination of communicable diseases such as COVID-19. Despite current scientific progress, many uses of gene editing of DNA sequences are ethically questionable, bordering upon antithetical to principles of human rights protections. The history of genetics, genetic modification of DNA sequences, and human rights has included pseudoscientific practices such as those of the eugenics movement, which dominated much of the nineteenth and early twentieth centuries. Later, United Nations measures such as the International Bill of Human Rights and the Universal Declaration on the Human Genome and Human Rights prompted improvements in human rights practices. Still, genetics rights and human rights in many nations required codification, as human rights standards—including the right to privacy and the protection of genetic data—have been challenged by a host of stakeholders and commercial entities. Ethical limits and international human rights protections relevant to the use of gene editing technology must be preserved. Considerable progress in disease diagnostics and therapeutics using gene editing and genetic engineering has bolstered the fight for the concurrent preservation of gene editing, ethics, and international human rights protections. As such, this chapter will examine the critical importance of upholding human rights as an essential component of the advancement of gene editing.

For thousands of years, humans have felt the need to understand the world around them—and ultimately manipulate it to best serve their needs. There are always ethical questions to address, especially when the manipulation involves the human genome. There is currently an urgent need to actively pursue those conversations as com- mercial gene sequencing and editing technologies have become more accessible and affordable. Gene Editing has enormous potential both as a research tool and a therapeutic intervention. While other types of gene editing are relatively uncontroversial, gene editing has been strongly resisted.

The farming of livestock has a critical role to play in global nutritional security and poverty alleviation. To meet these goals through more efficient, environmentally sustainable and animal welfare focused means, gene editing technologies could be integrated into current breeding programs. A common issue with gene editing in livestock zygotes is the high incidence of genetic mosaicism. Genetic mosaicism, characterised by a single individual carrying distinct genotypes in different cell lineages, can lead to inconsistent presentation of a desired trait phenotypically, or the absence of the intended genotype in the animal’s germline. This review explores the present status of genetic mosaicism associated with CRISPR-Cas9 gene editing in cattle, sheep, and pigs, and identifies four areas for refinement; (1) the type of CRISPR-Cas9 genome editor used; (2) the CRISPR-Cas9 formats and timing of gene editing during embryonic development; (3) the method used to deliver the genome editor and (4) the genetic screening strategies applied. We also discuss alternatives to direct zygote gene editing, including surrogate sire technology and blastomere separation, which circumvent the production of mosaic offspring. By exploring these avenues for reducing mosaicism, gene editing protocols in livestock could become more efficient and effective, which will ultimately pave the way for traits to be introduced that improve animal welfare standards and help address gaps in the security of global nutrition access.

Researchers views about perceived harms and benefits of gene editing: A study from the MENA region

The crisis of antibiotic resistance caused by the overuse of antibiotics has become a major threat to global agriculture and public health, and traditional livestock strategies have demonstrated significant limitations in addressing this challenge. Gene editing technologies, in particular the CRISPR-Cas9 system, offer innovative solutions to the problem of antibiotic resistance by regulating the gut microbiota of livestock and directly interfering with resistance genes. This study systematically explores cross-cutting applications of gene editing technology in livestock health management, focusing on three major themes: (1) Editing host genes to optimize gut microbiota balance; (2) Directly editing resistant genes of pathogenic bacteria to restore their sensitivity to antibiotics; (3) Developing collaborative strategies (phage delivery systems and probiotic engineering). The results showed that pigs with CD163 gene knocked out by CRISPR-Cas9 had complete resistance to porcine reproductive and respiratory syndrome. Engineered probiotics can remove 99.9% of resistant bacteria; The bifunctional delivery strategy reduced the intestinal drug-resistant bacterial load to <100 CFU/g. In addition, multi-omics techniques provide a precise tool for dynamically assessing the effects of gene editing on microbiome function. Conclusion: Through the new strategy of "host-microbiome co-editing", gene editing technology has the potential to be transformative in livestock microbiome regulation and drug resistance management, but challenges such as delivery efficiency, ecological risks and ethical controversies still need to be addressed, and its widespread application depends on technological innovation, interdisciplinary cooperation and the coordinated development of an international regulatory framework. To achieve the dual goals of agricultural sustainability and public health security.

Gene editing technology involves the modification of a specific target gene to obtain a new function or phenotype. Recent advances in clustered regularly interspaced short palindromic repeats (CRISPR)-Cas-mediated technologies have provided an efficient tool for genetic engineering of cells and organisms. Here, we review the three emerging gene editing tools (ZFNs, TALENs, and CRISPR-Cas) and briefly introduce the principle, classification, and mechanisms of the CRISPR-Cas systems. Strategies for gene editing based on endogenous and exogenous CRISPR-Cas systems, as well as the novel base editor (BE), prime editor (PE), and CRISPR-associated transposase (CAST) technologies, are described in detail. In addition, we summarize recent developments in the application of CRISPR-based gene editing tools for industrial microorganism and probiotics modifications. Finally, the potential challenges and future perspectives of CRISPR-based gene editing tools are discussed.

Gene editing technologies are playing an increasingly important role in the study of insect gene functions. Appropriate incubation humidity helps improve the hatching rate of insect eggs. However, to date, no study has evaluated the impact of humidity on the hatching rate of gene-edited eggs or the subsequent development of the larvae. Using the cotton bollworm Helicoverpa armigera (Lepidoptera: Noctuidae) as a model, we investigated the weight and hatching rate of microinjected eggs under different humidity conditions. We determined the larval development time and pupal weight and calculated the larval gene editing efficiency. The results showed that post-microinjection incubation under high humidity (RH = 95%) had a significant influence on CRISPR/Cas9 gene-edited eggs. The key research results indicate that, compared with the incubation environment with RH ≤ 80%, after 48 h of incubation under 95% RH, the hatching rate of H. armigera eggs increased by more than 27.5%, with higher egg saturation and greater egg weight; meanwhile, the larval developmental duration was shorter and the pupal weight was also greater. These results establish a humidity-regulated developmental recovery protocol for gene-edited insects, which provides theoretical and practical support for optimizing post-microinjection recovery processes in gene-edited insects.

Gene editing (GE) technologies are rapidly gaining traction as an alternative to genetically modified organisms (GMOs) in agriculture. While proponents claim the critical need for GE to address climate change and food security and assert its similarity to conventional breeding, critics argue that these technologies bring similar concerns to GMOs, such as supporting industrial agriculture and enhancing corporate control and ownership. But how do public groups make sense of these technologies? While incorporating public concerns is key to responsible and ethical innovation, minimal research explores how people make sense of emerging applications. We offer an exploratory Q study that investigates how one public group applies interpretive frames to understand applications of novel GE and related technologies. We find participants apply three different frames, invoking applications as (1) necessitating a system critical lens, (2) worthy of pragmatic of consideration, or (3) a deeply ambiguous prospect. These frames, we argue, articulate visions of particular sociotechnical futures, most of which are contrary or orthogonal to proponents’ assumptions. Instead, we find participants concerned less with the precision of techniques or the origin of genes used and more often with whether these applications reify dominant industrial practices and if viable alternatives exist.

Biotechnology has made significant advances in recent years, and the area of genetic engineering is progressing day by day, generating several advantages. Through the new ability to precisely change and modify the genomes of living organisms, genome editing technology has transformed genetic and biological research. Genome editing technology first appeared in the 1990s, and different approaches for targeted gene editing have subsequently been created. The fields of functional genomics and crop improvement have been transformed by advances in genome editing tools. CRISPR/Cas9 (clustered regularly interspaced short palindromic repeat)-Cas9 is a versatile genetic engineering tool based on the complementarity of the guide RNA (gRNA) to a specific sequence and the endonuclease activity of the Cas9 endonuclease. This RNA-guided genome editing tool has produced variations in plant biology fields. CRISPR technology is continually improving, allowing for more genetic manipulations such as creating knockouts, precise changes, and targeted gene activation and repression. Soybean is a leguminous crop, high in protein and oil contents that are used for poultry and livestock feed industry. In this chapter, we focus on the recent advances in CRISPR/Cas9-based gene editing technology and discuss the challenges and opportunities to harnessing this innovative technology for targeted improvement of traits in soybean and other crops.

With the rapid population growth, there is an urgent need for innovative crop improvement approaches to meet the increasing demand for food. Classical crop improvement approaches involve, however, a backbreaking process that cannot equipoise with increasing crop demand. RNA-based approaches i.e., RNAi-mediated gene regulation and the site-specific nuclease-based CRISPR/Cas9 system for gene editing has made advances in the efficient targeted modification in many crops for the higher yield and resistance to diseases and different stresses. In functional genomics, RNA interference (RNAi) is a propitious gene regulatory approach that plays a significant role in crop improvement by permitting the downregulation of gene expression by small molecules of interfering RNA without affecting the expression of other genes. Gene editing technologies viz. the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein (CRISPR/Cas) have appeared prominently as a powerful tool for precise targeted modification of nearly all crops’ genome sequences to generate variation and accelerate breeding efforts. In this regard, the review highlights the diverse roles and applications of RNAi and CRISPR/Cas9 system as powerful technologies to improve agronomically important plants to enhance crop yields and increase tolerance to environmental stress (biotic or abiotic). Ultimately, these technologies can prove to be important in view of global food security and sustainable agriculture.

Hematopoietic stem cells (HSCs) are rare cells, which housed in the adult bone marrow. They maintain all types of differentiated blood cells throughout life. Due to limited availability of HSCs for transplantation, treatment of various inherited bone marrow disorders and anemia requires the development of HSC expansion and gene editing technologies. To this end, various studies addressed the use of cytokines and growth factors for HSC expansion. Major hurdle with these studies was found to be spontaneous differentiation of HSCs into different lineages during ex vivo procedure. In addition, cost efficient approaches were needed. Thus, studies move on to the identification of small molecules and development of RNA interference technologies with potential to enhance cell cycle progression and block inhibitory signaling mechanisms during ex vivo HSC expansion as well as single cell expansion of HSCs following gene editing studies. This review aims to highlight developments in hematopoietic stem cells expansion and gene editing technologies.

In recent years, CRISPR/Cas9 gene editing technology has emerged as a powerful genetic tool with potential application in aquaculture. Crassostrea gigas, as a valuable species in aquaculture, holds promising potential for genetic enhancement and breeding through gene editing. However, the lack of efficient promoters for driving exogenous gene expression poses a major obstacle in bivalve gene editing. In this study, we isolated the promoter sequences of the β-tub and histone H3.3A genes from C. gigas. DNA expression constructs were generated by linking the promoters with the enhanced green fluorescent protein (EGFP) reporter and compared with the promoter activity of the endogenous EF-1α gene and an exogenous OsHV-1 promoter in C. gigas embryos. All four promoters effectively drive the expression of EGFP during early embryonic development in C. gigas. Among these four promoters, the β-tub promoter is the most potent promoter in driving EGFP expression in C. gigas embryos as early as 4.5h after fertilization. The OsHV-1 promoter showed similar activity as β-tub promoter and appeared to be more active than the EF-1α and histone H3.3A promoters in C. gigas embryos. Furthermore, we assessed their performance in other three C. gigas relatives (Crassostrea ariakensis, Crassostrea nippona, and Crassostrea sikamea) and similar results were found. Collectively, these data suggest that the β-tub promoter is an effective promoter in directing gene expression in directing gene expression in oyster embryos, thus offering a potential application for gene editing in bivalves.

Gene editing technologies have revolutionized therapeutic development, offering potentially curative and preventative strategies for cardiovascular disease (CVD), which remains a leading global cause of morbidity and mortality. This review provides an introduction to the state-of-the-art gene editing tools-including ZFNs, TALENs, CRISPR/Cas9 systems, base editors, and prime editors-and evaluates their application in lipid metabolic pathways central to CVD pathogenesis. Emphasis is placed on targets such as PCSK9, ANGPTL3, CETP, APOC3, ASGR1, LPA, and IDOL, supported by findings from human genetics, preclinical models, and recent first-in-human trials. Emerging delivery vehicles (AAVs, LNPs, lentivirus, virus-like particles) and their translational implications are discussed. The review highlights ongoing clinical trials employing liver-targeted in vivo editing modalities (LivGETx-CVD) and provides insights into challenges in delivery, off-target effects, genotoxicity, and immunogenicity. Collectively, this review captures the rapid progress of LivGETx-CVD from conceptual innovation to clinical application, and positions gene editing as a transformative, single-dose strategy with the potential to redefine prevention and long-term management of dyslipidemia and atherosclerotic cardiovascular disease.

The rapid growth of the field of gene editing can largely be attributed to the discovery and optimization of designer endonucleases. These include zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regular interspersed short palindromic repeat (CRISPR) systems including Cas9, Cas12a, and structure-guided nucleases. Zebrafish (Danio rerio) have proven to be a powerful model system for genome engineering testing and applications due to their external development, high fecundity, and ease of housing. As the zebrafish gene editing toolkit continues to grow, it is becoming increasingly important to understand when and how to utilize which of these technologies for maximum efficacy in a particular project. While CRISPR-Cas9 has brought broad attention to the field of genome engineering in recent years, designer endonucleases have been utilized in genome engineering for more than two decades. This chapter provides a brief overview of designer endonuclease and other gene editing technologies in zebrafish as well as some of their known functional benefits and limitations depending on specific project goals. Finally, selected prospects for additional gene editing tools are presented, promising additional options for directed genomic programming of this versatile animal model system.