Human Genome Editing Research Articles

Human Genome Editing: Managing Technological Risks through Legal Means

Objective: to determine theoretical approaches to the legal regulation of reprogenetic editing, taking into account the risk-oriented approach and the practice of regulation of such breakthrough technologies in different jurisdictions; to outline further regulatory and managerial steps to be taken for the technology development.Methods: general scientific methods of analysis and synthesis, classification, system and functional approaches; specific scientific methods: formal-legal, comparative-legal, and historical-legal.Results: the research shows the possible approaches to the regulation of genetic editing for reproductive purposes. The considered variants are evaluated from the viewpoint of risk-oriented approach; conditions and peculiarities of various regulatory mechanisms’ application are determined; the current Russian regulation in this sphere is assessed. The analysis allows concluding that the prohibition or significant restriction of the developing technology of reprogenetic editing has no irrefutable grounds. Moreover, it may lead to the results opposite to those declared by its proponents. In this regard, it is necessary to develop the discussion in a constructive and iterative way and involve all stakeholders in it, including the scientific community.

Will South Africa become first country to accept controversial form of human genome editing?

Will South Africa become first country to accept controversial form of human genome editing?

Genetically humanized phenylketonuria mouse model as a testing tool for human genome editing in fertilized eggs.

Targeted genome editing has made significant advancements; however, safety and ethical issues have not been fully elucidated, resulting in strict control of the technique. We tested genome editing tools on gametes from a genetically humanized mouse model using a phenylketonuria (PKU) mouse model to gain insights into genome editing in human embryos. The human PKU mouse model PahhR111X mice was generated. The junctional region between exon 3 and intron 3 of Pah was replaced with a 120 bp corresponding human PAH sequence, including the pathogenic common variant c.331C > T in PahhR111X mice. PahhR111X mice successfully recapitulated the PKU phenotype and showed cognitive dysfunction and depressive-like behavior, which are observed in human patients with PKU. Genome editing was applied to fertilized eggs of PahhR111X mice utilizing sgRNA that targets the human sequence. Mice with the corrected allele exhibited normal serum phenylalanine levels. Through genome editing, we validated the utility of sgRNA. The genetically humanized mouse model suggested that germ-line genome editing of the pathogenic variant may be feasible for monogenic disorders by revealing the recovery of the phenotype; however, there are remaining issues with the tool, including its efficiency and accuracy. This genome editing protocol using a genetically humanized mouse model will provide insights for improving current issues and contribute to the establishment of heritable human genome editing protocols.

Before We Forget its Horrors: When Does Human Genome Editing Not Become Eugenics?

Before We Forget its Horrors: When Does Human Genome Editing Not Become Eugenics?

Limits of Interference into the Human Genome

Introduction: The use of advances in genomics in practical medicine can positively affect the quality of services in the healthcare sector, as well as the results of applied medical research. Objectives: The purpose of the article is to research regulatory and scientific approaches to the concept of the human genome as an object of legal relations, as well as to determine the limits of interference into a human being’s genomic integrity. Materials and methods: Regarding the chosen subject matter of research, the author while preparing the scientific article has used materials of international regulatory legal acts, Ukrainian and foreign legal literature. To achieve the set aim, the author has used general scientific and special methods of scientific cognition. These methods combined with the scientific principles of objectivity, scientificity and reasonableness, have provided a comprehensive research of the limits of interference into the human genome. Results: The concept of the human genome, as well as its material and non-material components, have been studied from a biological and legal point of view. The private and legal nature of the human genome as a personal non-property good has been defined. The concept of the individual’s integrity, enshrined in international regulatory legal acts, has been analyzed from the point of view of its coverage of a human being’s genomic integrity. It has been established that a violation of genomic integrity is possible both by the intentional actions of third parties and as a result of genome changes due to the action of natural factors. The concepts and types of the human genome editing have been defined. The potential reasons for banning the editing of human germline cells have been critically analyzed. The legal nature of interference into the human genome from the point of view of eugenical practices has been studied. The author has studied the possibility of classifying the use of vaccines on the mRNA platform as interferences into the human genome. Conclusions: It has been concluded that there is a need to recognize the human genome as a personal non-property good, and, accordingly, as an object of private rights. The author has stated own point of view on the inadmissibility of arbitrary interference into human genomic integrity and the existing types of interference into the human genome.

Human heritable genome editing and its governance: views of scientists and governance professionals

Heritable human genome editing has garnered significant attention in scholarly and lay media, yet questions remain about whether, when, and how heritable genome editing ought to proceed. Drawing on interviews with scientists who use genome editing in their research and professionals engaged in human genome editing governance efforts, we examine their views on the permissibility of heritable genome editing and the governance strategies they see as necessary and realistic. For both issues, we found divergent views from respondents. We place the views of these scientists and governance professionals within the context of the larger bioethical discussion of heritable genome editing governance, along a continuum of hard to soft approaches. These respondents’ views highlight the challenges of various hard forms of governance and the potential virtues of soft governance approaches.

Structure-guided engineering of type I-F CASTs for targeted gene insertion in human cells.

Conventional genome editing tools rely on DNA double-strand breaks (DSBs) and host recombination proteins to achieve large insertions, resulting in a heterogeneous mixture of undesirable editing outcomes. We recently leveraged a type I-F CRISPR-associated transposase (CAST) from the Pseudoalteromonas Tn7016 transposon (PseCAST) for DSB-free, RNA-guided DNA integration in human cells, taking advantage of its programmability and large payload capacity. PseCAST is the only characterized CAST system that has achieved human genomic DNA insertions, but multiple lines of evidence suggest that DNA binding may be a critical bottleneck that limits high-efficiency activity. Here we report structural determinants of target DNA recognition by the PseCAST QCascade complex using single-particle cryogenic electron microscopy (cryoEM), which revealed novel subtype-specific interactions and RNA-DNA heteroduplex features. By combining our structural data with target DNA library screens and rationally engineered protein mutations, we uncovered CAST variants that exhibit increased integration efficiency and modified PAM stringency. Structure predictions of key interfaces in the transpososome holoenzyme also revealed opportunities for the design of hybrid CASTs, which we leveraged to build chimeric systems that combine high-activity DNA binding and DNA integration modules. Collectively, our work provides unique structural insights into type I-F CAST systems while showcasing multiple diverse strategies to investigate and engineer new RNA-guided transposase architectures for human genome editing applications.

Changing the regulation of human germline genome editing: what does a truly broad societal debate entail?

ABSTRACT How gene editing technologies should be regulated and whether Human Germline Genome Editing (HGGE) should become lawful remain very divisive questions in the legal, political and bioethical fields. At the same time, there is universal agreement that any changes in legislation should be preceded, or accompanied, by broad public deliberation. What does, however, a truly broad societal debate on HGGE regulation actually entail? In this paper, we focus on this issue and articulate a specific understanding of broadness for societal debates on the regulation of technological developments. We then apply it to the context of HGGE and show that a truly broad societal debate on its regulation would require reflecting: (1) on the significance of public preferences regarding genetically-related children for law and policy; and (2) on the potential of HGGE as a tool for global disease burden reduction. We conclude by outlining the implications for policymakers and further research.

Moral Nuances in Broad Policies

AbstractIn the September‐October 2024 issue of the Hastings Center Report, two pieces examine attitudes toward and policy on medical aid in dying (MAID). An essay by Anna Elsner and colleagues analyzes terminology, including euphemistic language, used in Canada and other countries to refer to this practice. The authors recommend explicit public discussion of the values at stake in the use of this terminology. An article by Em Walsh concerns a subset of people who could become eligible for MAID under Canada's proposed expansion of eligibility for this assistance: people suffering from poverty‐induced depression. Cautioning that the expansion of PAD could exacerbate inequality, Walsh offers six recommendations for policy‐makers’ consideration. The issue's lead article, by Eric Juengst and colleagues, focuses on governance issues that could be raised by human genome editing research that aims to strengthen individuals’ resistance to disease beyond what is regarded as the human functional range. Juengst et al. identify and analyze three potential principles that could help policy‐makers navigate what can be a blurry line between goals of prevention and enhancement in human genome editing research.

High Throughput Variant Libraries and Machine Learning Yield Design Rules for Retron Gene Editors.

The bacterial retron reverse transcriptase system has served as an intracellular factory for single-stranded DNA in many biotechnological applications. In these technologies, a natural retron non-coding RNA (ncRNA) is modified to encode a template for the production of custom DNA sequences by reverse transcription. The efficiency of reverse transcription is a major limiting step for retron technologies, but we lack systematic knowledge of how to improve or maintain reverse transcription efficiency while changing the retron sequence for custom DNA production. Here, we test thousands of different modifications to the retron-Eco1 ncRNA and measure DNA production in pooled variant library experiments, identifying regions of the ncRNA that are tolerant and intolerant to modification. We apply this new information to a specific application: the use of the retron to produce a precise genome editing donor in combination with a CRISPR-Cas9 RNA-guided nuclease (an editron). We use high-throughput libraries in S. cerevisiae to additionally define design rules for editrons. We extend our new knowledge of retron DNA production and editron design rules to human genome editing to achieve the highest efficiency retron-Eco1 editrons to date.

Challenging the Boundaries Between Treatment, Prevention, and Enhancement in Human Genome Editing.

Traditional distinctions between treatment and enhancement goals for human genome editing (HGE) have animated oversight considerations, yet these categories have been complicated by the addition of prevention as a possible target for HGE applications. To assess the role these three categories might play in continued HGE governance efforts, we report on interviews with genome editing scientists and governance group members. While some accepted traditional distinctions between treatment and enhancement and rejected the latter as unacceptable, others argued that the concept of enhancement is largely irrelevant or not as morally problematic as suggested. Others described how preventive goals for HGE create gray zones where prevention and enhancement may be difficult to distinguish, which may stymie uses of HGE. We conclude by discussing the governance implications of these various understandings of treatment, prevention, and enhancement as HGE research moves beyond the treatment of serious disease to embrace longer range preventive goals.

Assessing and engineering the IscB-ωRNA system for programmed genome editing.

OMEGA RNA (ωRNA)-guided endonuclease IscB, the evolutionary ancestor of Cas9, is an attractive system for in vivo genome editing because of its compact size and mechanistic resemblance to Cas9. However, wild-type IscB-ωRNA systems show limited activity in human cells. Here we report enhanced OgeuIscB, which, with eight amino acid substitutions, displayed a fourfold increase in in vitro DNA-binding affinity and a 30.4-fold improvement in insertion-deletion (indel) formation efficiency in human cells. Paired with structure-guided ωRNA engineering, the enhanced OgeuIscB-ωRNA systems efficiently edited the human genome across 26 target sites, attaining up to 87.3% indel and 62.2% base-editing frequencies. Both wild-type and engineered OgeuIscB-ωRNA showed moderate fidelity in editing the human genome, with off-target profiles revealing key determinants of target selection including an NARR target-adjacent motif (TAM) and the TAM-proximal 14 nucleotides in the R-loop. Collectively, our engineered OgeuIscB-ωRNA systems are programmable, potent and sufficiently specific for human genome editing.

Anticipatory gaps challenge the public governance of heritable human genome editing

Considering public moral attitudes is a hallmark of the anticipatory governance of emerging biotechnologies, such as heritable human genome editing. However, such anticipatory governance often overlooks that future morality is...

The Problems of Human Embryos Genome Editing from the Position of Islam Denominations

Biomedical technology is one of the most relevant and rapidly developing branches of science. In response to the major problems of bioethics and bio-law, bioethical dilemmas emerge in society, which constrain the abuse of new technologies. Medical discoveries, on the one hand, can greatly facilitate the life of humankind, but, on the other hand, the problem of interference in human nature actualizes the most fundamental questions concerning its ontology, the boundaries of permissible transformations, the responsibility of a scientist and a specialist, applying the latest technologies, for remote and unpredictable consequences, due to the integrity and interconnectedness of various aspects of human nature. This paper presents the experience of generalizing the attitude of the main Islamic confessions and different approaches in the legislation of Islamic countries to the problem of editing the human embryo genome. Based on a review of scientific and religious literature, it is concluded that, although the Islamic world is increasingly using Western models of behavior, in matters of the permissibility of editing the human embryo genome from the point of view of Islamic bioethics, it is necessary to rely on the principles of Sharia and multidisciplinary knowledge.

Library-Assisted Evolution in Eukaryotic Cells Yield Adenine Base Editors with Enhanced Editing Specificity.

The current-generation adenine base editor (ABE) ABE8e, which has evolved from the prokaryotic evolution system, exhibits high efficiency in mediating A-to-Gconversion and is presumed to be promising for gene therapy. However, its much wider editing window and substantially higher off-target editing activity restricted its applications in precise base editing for therapeutic use. This study uses a library-assistedprotein evolution approach using eukaryotic cells to generate ABE variants with improved specificity and reduced off-target editing while maintaining high activity in human cells. The study generated an expanded set of ABEs with efficient editing activities and chose fourevolved variants thatoffered either similar or modestly higher efficiency within a narrower editing window of protospacer position ≈4-7 compared to that of ABE8e in human cells, which would enable minimized bystander editing. Moreover, these variants resulted in reduced off-target editing events when delivered as plasmid or mRNA into human cells. Finally, these variants can install both disease-suppressing mutations and disease-correcting mutations efficiently with minimal undesired bystander editing making them promising approaches for specific therapeutic edits. In summary, the work establishes a mutant-library-assisted protein evolution method in eukaryotic cells and generates alternative ABE variants as efficient tools for precise human genome editing.

Overcoming vulnerability by editing the germline?

The concept of vulnerability has become widely acknowledged as a fundamental concept for medical ethics and research ethics, yet rarely considered with respect to ethical assessments of human germline genome editing. A first aim of this paper is to make vulnerability ethics considerations fruitful for issues related to these technical innovations. The possibility of altering the genome promises to overcome forms of vulnerability inherently connected to our existence as physical beings and would hence allow to increase the resilience of human nature or even to move evolution forward by equipping people with new character traits and enhanced capabilities. I suggest a more fine-grained distinction of various applications purposes than the dichotomy of therapeutic and enhancement. I support the rejection of most application purposes as ‘therapeutic’ and claim that framing them as ‘therapeutic’ in the context of the current discursive constellation runs the risk of accentuating existing vulnerabilities. With respect to intergenerational responsibilities, I reject the view that editing the germline necessarily leads to corrupt intergenerational relations based on which it must be categorically excluded. I conclude that it is nevertheless important to take a very close look at the challenges that arise, especially from a vulnerability perspective, before irreversible facts are created overhastily.

Regulatory safeguards needed if preimplantation genetic testing for polygenic risk scores (PGT-P) is permitted in Singapore

Singapore, a highly affluent island city-state located in Southeast Asia, has increasingly leveraged new assisted reproductive technologies (ART) to overcome its dismal fertility rates in recent years. A new frontier...

Governing with public engagement: an anticipatory approach to human genome editing.

In response to calls for public engagement on human genome editing (HGE), which intensified after the 2018 He Jiankui scandal that resulted in the implantation of genetically modified embryos, we detail an anticipatory approach to the governance of HGE. By soliciting multidisciplinary experts' input on the drivers and uncertainties of HGE development, we developed a set of plausible future scenarios to ascertain publics values-specifically, their hopes and concerns regarding the novel technology and its applications. In turn, we gathered a subset of multidisciplinary experts to propose governance recommendations for HGE that incorporate identified publics' values. These recommendations include: (1) continued participatory public engagement; (2) international harmonization and transparency of multiple governance levers such as professional and scientific societies, funders, and regulators; and (3) development of a formal whistleblower framework.

Heritable human genome editing: correction, selection and treatment.

Heritable human genome editing (HHGE) to correct a nuclear gene sequence that would result in a serious genetic condition in a future child is presented as 'treatment' in various ethics and policy materials, and as morally preferable to the 'selection' practice of preimplantation genetic testing (PGT), which is subject to the disability critique. However, whether HHGE is 'treatment' for a future child, or another form of 'selection', or whether HHGE instead 'treats' prospective parents, are now central questions in the debate regarding its possible legalisation. This article argues that the idea of 'treatment' for a future child is largely a proxy for 'seriousness of purpose', intended to distinguish HHGE to avoid serious genetic conditions from less obviously justifiable uses; that HHGE is best understood, and morally justified, as a form of 'treatment' for prospective parents who strongly desire an unaffected genetically related child and who have no, or poor, options to achieve this; that HHGE would be morally permissible if consistent with that child's welfare; that legalisation is supportable with reference to the right to respect for private and family life under Article 8 of the European Convention on Human Rights; and that HHGE is morally distinguishable from PGT.

Recent Developments in the Regulation of Heritable Human Genome Editing

In 2018, the Chinese scientist He Jiankui presented his research at the Second International Summit on Human Genome Editing in Hong Kong. While it was intended that he facilitate a workshop, he was instead called on to present his research in heritable human genome editing, where he made the announcement that he had taken great strides in advancement of his research, to the extent that he had gene-edited human embryos and that this had resulted in the live births of two children. While his research ethic and methodology was interrogated, he insisted that two children, twin girls, had been born healthy and that there was another pregnancy (at the time) where birth of a third gene edited child would be imminent. This announcement generated a ripple effect in the scientific community and exposed the gaps in regulation and absence of law relating to the technology. This resulted in a flurry of activity and conversation around regulation of the technology, which scientists stated was not ready for human trials. This article reviews the Third Summit which was held in London in March 2023 and comments on the latest developments in the regulation of heritable human genome editing.