RESUMO
Gene editing technologies hold promise for enabling the next generation of adoptive cellular therapies. In conventional gene editing platforms that rely on nuclease activity, such as clustered regularly interspaced short palindromic repeats CRISPR-associated protein 9 (CRISPR-Cas9), allow efficient introduction of genetic modifications; however, these modifications occur via the generation of DNA double-strand breaks (DSBs) and can lead to unwanted genomic alterations and genotoxicity. Here, we apply a novel modular RNA aptamer-mediated Pin-point base editing platform to simultaneously introduce multiple gene knockouts and site-specific integration of a transgene in human primary T cells. We demonstrate high editing efficiency and purity at all target sites and significantly reduced frequency of chromosomal translocations compared with the conventional CRISPR-Cas9 system. Site-specific knockin of a chimeric antigen receptor and multiplex gene knockout are achieved within a single intervention and without the requirement for additional sequence-targeting components. The ability to perform complex genome editing efficiently and precisely highlights the potential of the Pin-point platform for application in a range of advanced cell therapies.
Assuntos
Aptâmeros de Nucleotídeos , Sistemas CRISPR-Cas , Edição de Genes , Técnicas de Inativação de Genes , Linfócitos T , Humanos , Edição de Genes/métodos , Aptâmeros de Nucleotídeos/genética , Linfócitos T/metabolismo , Linfócitos T/imunologia , Imunoterapia Adotiva/métodos , Receptores de Antígenos Quiméricos/genética , Receptores de Antígenos Quiméricos/metabolismo , Técnicas de Introdução de Genes/métodos , TransgenesRESUMO
Environmental injustice refers to the unequal burden of pollutants on groups with lower socioeconomic status. An increasing number of studies have identified associations between high levels of pollution and socioeconomic disadvantage. However, few studies have controlled adequately for spatio-temporal variations in pollution. This study uses a Bayesian approach to explore the association between socioeconomic disadvantage and pollution in Mexico City Metropolitan Area. We quantify the association of socioeconomic disadvantage with PM10 and ozone and evaluate the impact of accounting for spatio-temporal structure of the pollution data. We find a significant positive association between socio-economic disadvantage and pollution for levels of PM10, but not ozone. The inclusion of the spatio-temporal element in the modeling results in improved weaker estimates of this association but this does not alter results substantially. These findings confirm the robustness of previous studies that found signs of environmental injustice where spatio-temporal variations have not been explicitly considered, confirming that targeted policies to reduce pollution in socio-economically disadvantaged areas are required.
Assuntos
Poluentes Atmosféricos , Poluição do Ar , Ozônio , Teorema de Bayes , Poluentes Atmosféricos/análise , México , Poluição do Ar/análise , Ozônio/análise , Fatores Socioeconômicos , Material Particulado/análiseRESUMO
Streptococcus pyogenes Cas9 (SpCas9) and derived enzymes are widely used as genome editors, but their promiscuous nuclease activity often induces undesired mutations and chromosomal rearrangements. Several strategies for mapping off-target effects have emerged, but they suffer from limited sensitivity. To increase the detection sensitivity, we develop an off-target assessment workflow that uses Duplex Sequencing. The strategy increases sensitivity by one order of magnitude, identifying previously unknown SpCas9's off-target mutations in the humanized PCSK9 mouse model. To reduce off-target risks, we perform a bioinformatic search and identify a high-fidelity Cas9 variant of the II-B subfamily from Parasutterella secunda (PsCas9). PsCas9 shows improved specificity as compared to SpCas9 across multiple tested sites, both in vitro and in vivo, including the PCSK9 site. In the future, while PsCas9 will offer an alternative to SpCas9 for research and clinical use, the Duplex Sequencing workflow will enable a more sensitive assessment of Cas9 editing outcomes.
Assuntos
Pró-Proteína Convertase 9 , Translocação Genética , Animais , Camundongos , Pró-Proteína Convertase 9/genética , Sistemas CRISPR-Cas/genética , Mutação , Endonucleases/genética , Streptococcus pyogenes/genéticaRESUMO
In southern Europe, land abandonment and an unbalanced investment toward fire suppression instead of prevention has gradually increased wildfire risk, which calls for a paradigm change in fire management policies. Here we combined scenario analysis, fire landscape modelling, and economic tools to identify which land-use policies would reduce the expected wildfire-related losses in the Transboundary Biosphere Reserve 'Gerês-Xurés' (Spain-Portugal). To do so, we applied the least-cost-plus-net-value-change approach and estimated net changes in wildfire damages based on their implications for the 2010-2050 period and five ecosystem services: agriculture, pasture, timber, recreation and climate regulation. Four land-use scenarios were considered: (1) Business as Usual (BAU); (2) fire-smart, fostering more fire-resistant (less flammable) and/or fire-resilient landscapes (fire-smart); (3) High Nature Value farmlands (HNVf), wherein the abandonment of extensive agriculture is reversed; and (4) a combination of HNVf and fire-smart. HNVf is the best scenario for suppression cost savings, but it generates the lowest net present value of societal benefits from climate regulation. In fact, the most efficient scenario with the lowest societal discounted net suppression costs and change on ecosystem services damages is the HNVf + fire-smart scenario, as it also generates suppression cost savings from agricultural expansion, and lead to a significant reduction in damages on timber and recreational benefits. Therefore, reverting land abandonment through recultivation and promoting fire-resistant tree species is the most efficient way to reduce wildfire hazard. In this sense, payments for ecosystem services should reward farmers and landowners for their role in wildfire prevention. This study improves the understanding of the financial and societal benefits derived from reducing fire suppression spending and ecosystem services damage by undertaking fire-smart land-use strategies, which can be essential to enhance local stakeholders' support for Payments of Ecosystem Services policies for wildfire prevention.
Assuntos
Incêndios , Incêndios Florestais , Ecossistema , Espanha , Agricultura , Conservação dos Recursos NaturaisRESUMO
Cumulative and synergistic impacts from environmental pressures, particularly in low-lying tropical coastal regions, present challenges for the governance of ecosystems, which provide natural resource-based livelihoods for communities. Here, we seek to understand the relationship between responses to the impacts of El Niño and La Niña events and the vulnerability of mangrove-dependent communities in the Caribbean region of Colombia. Using two case study sites, we show how communities are impacted by, and undertake reactive short-term responses to, El Niño and La Niña events, and how such responses can affect their adaptive capacity to progressive environmental deterioration. We show that certain coping measures to climate variability currently deliver maladaptive outcomes, resulting in circumstances that could contribute to system 'lock-in' and engender undesirable ecological states, exacerbating future livelihood vulnerabilities. We highlight the significant role of social barriers on vulnerabilities within the region, including perceptions of state abandonment, mistrust and conflicts with authorities. Opportunities to reduce vulnerability include enhancing the communities' capacity to adopt more positive and preventative responses based on demonstrable experiential learning capacity. However, these will require close cooperation between formal and informal organisations at different levels, and the development of shared coherent adaptation strategies to manage the complexity of multiple interacting environmental and climatic pressures.
Assuntos
Mudança Climática , Ecossistema , Região do Caribe , Colômbia , Emprego , Previsões , HumanosRESUMO
The number of alien plants escaping from cultivation into native ecosystems is increasing steadily. We provide an overview of the historical, contemporary and potential future roles of ornamental horticulture in plant invasions. We show that currently at least 75% and 93% of the global naturalised alien flora is grown in domestic and botanical gardens, respectively. Species grown in gardens also have a larger naturalised range than those that are not. After the Middle Ages, particularly in the 18th and 19th centuries, a global trade network in plants emerged. Since then, cultivated alien species also started to appear in the wild more frequently than non-cultivated aliens globally, particularly during the 19th century. Horticulture still plays a prominent role in current plant introduction, and the monetary value of live-plant imports in different parts of the world is steadily increasing. Historically, botanical gardens - an important component of horticulture - played a major role in displaying, cultivating and distributing new plant discoveries. While the role of botanical gardens in the horticultural supply chain has declined, they are still a significant link, with one-third of institutions involved in retail-plant sales and horticultural research. However, botanical gardens have also become more dependent on commercial nurseries as plant sources, particularly in North America. Plants selected for ornamental purposes are not a random selection of the global flora, and some of the plant characteristics promoted through horticulture, such as fast growth, also promote invasion. Efforts to breed non-invasive plant cultivars are still rare. Socio-economical, technological, and environmental changes will lead to novel patterns of plant introductions and invasion opportunities for the species that are already cultivated. We describe the role that horticulture could play in mediating these changes. We identify current research challenges, and call for more research efforts on the past and current role of horticulture in plant invasions. This is required to develop science-based regulatory frameworks to prevent further plant invasions.
Assuntos
Jardinagem , Espécies Introduzidas , Plantas/classificação , Comércio , América do Norte , Dispersão VegetalRESUMO
The maintenance of livestock health depends on the combined actions of many different actors, both within and across different regulatory frameworks. Prior work recognised that private risk management choices have the ability to reduce the spread of infection to trading partners. We evaluate the efficiency of farmers' alternative biosecurity choices in terms of their own-benefits from unilateral strategies and quantify the impact they may have in filtering the disease externality of trade. We use bovine viral diarrhoea (BVD) in England and Scotland as a case study, since this provides an example of a situation where contrasting strategies for BVD management occur between selling and purchasing farms. We use an agent-based bioeconomic model to assess the payoff dependence of farmers connected by trade but using different BVD management strategies. We compare three disease management actions: test-cull, test-cull with vaccination and vaccination alone. For a two-farm trading situation, all actions carried out by the selling farm provide substantial benefits to the purchasing farm in terms of disease avoided, with the greatest benefit resulting from test-culling with vaccination on the selling farm. Likewise, unilateral disease strategies by purchasers can be effective in reducing disease risks created through trade. We conclude that regulation needs to balance the trade-off between private gains from those bearing the disease management costs and the positive spillover effects on others.
Assuntos
Criação de Animais Domésticos/métodos , Doença das Mucosas por Vírus da Diarreia Viral Bovina/prevenção & controle , Controle de Doenças Transmissíveis/métodos , Fazendeiros , Gestão de Riscos/métodos , Abate de Animais/economia , Abate de Animais/métodos , Criação de Animais Domésticos/economia , Animais , Doença das Mucosas por Vírus da Diarreia Viral Bovina/transmissão , Bovinos , Controle de Doenças Transmissíveis/economia , Efeitos Psicossociais da Doença , Humanos , Gado , Modelos Econômicos , Gestão de Riscos/economia , Índice de Gravidade de Doença , Reino Unido , VacinaçãoRESUMO
Alien species can have major ecological and socioeconomic impacts in their novel ranges and so effective management actions are needed. However, management can be contentious and create conflicts, especially when stakeholders who benefit from alien species are different from those who incur costs. Such conflicts of interests mean that management strategies can often not be implemented. There is, therefore, increasing interest in engaging stakeholders affected by alien species or by their management. Through a facilitated workshop and consultation process including academics and managers working on a variety of organisms and in different areas (urban and rural) and ecosystems (terrestrial and aquatic), we developed a framework for engaging stakeholders in the management of alien species. The proposed framework for stakeholder engagement consists of 12 steps: (1) identify stakeholders; (2) select key stakeholders for engagement; (3) explore key stakeholders' perceptions and develop initial aims for management; (4) engage key stakeholders in the development of a draft management strategy; (5) re-explore key stakeholders' perceptions and revise the aims of the strategy; (6) co-design general aims, management objectives and time frames with key stakeholders; (7) co-design a management strategy; (8) facilitate stakeholders' ownership of the strategy and adapt as required; and (9) implement the strategy and monitor management actions to evaluate the need for additional or future actions. In case additional management is needed after these actions take place, some extra steps should be taken: (10) identify any new stakeholders, benefits, and costs; (11) monitor engagement; and (12) revise management strategy. Overall, we believe that our framework provides an effective approach to minimize the impact of conflicts created by alien species management.
Assuntos
Ecologia , Ecossistema , Espécies IntroduzidasRESUMO
The ornamental plant trade has been identified as a key introduction pathway for plant pathogens. Establishing effective biosecurity measures to reduce the risk of plant pathogen outbreaks in the live plant trade is therefore important. Management of invasive pathogens has been identified as a weakest link public good, and thus is reliant on the actions of individual private agents. This paper therefore provides an analysis of the impact of the private agents' biosecurity decisions on pathogen prevention and control within the plant trade. We model the impact that an infectious disease has on a plant nursery under a constant pressure of potentially infected input plant materials, like seeds and saplings, where the spread of the disease reduces the value of mature plants. We explore six scenarios to understand the influence of three key bioeconomic parameters; the disease's basic reproductive number, the loss in value of a mature plant from acquiring an infection and the cost-effectiveness of restriction. The results characterise the disease dynamics within the nursery and explore the trade-offs and synergies between the optimal level of efforts on restriction strategies (actions to prevent buying infected inputs), and on removal of infected plants in the nursery. For diseases that can be easily controlled, restriction and removal are substitutable strategies. In contrast, for highly infectious diseases, restriction and removal are often found to be complementary, provided that restriction is cost-effective and the optimal level of removal is non-zero.
RESUMO
Ornamental horticulture has been recognized as the main pathway for plant invasions worldwide. We examined the link between propagule pressure created by the presence of ornamental plants in the market and their ability to escape from cultivation and establish in the wild. A random sample of 534 non-native ornamental species on sale in nineteenth century Britain showed that 27% of these species were recorded growing outside cultivation and 30% of those were established. Species that had escaped from cultivation were more frequently on sale both in the nineteenth century and today than nonescaping species. We used logit regression models to identify biological and socioeconomic variables that affect species' abilities to escape cultivation and become established. Frequencies in the market in the nineteenth century and today were good explanatory variables that distinguished escaping from nonescaping species, whereas for the transition from casual to established status these two socioeconomic variables were either absent or only of weak significance. Biological characteristics that increased the probability that a species would escape from cultivation were species height, a European native range, and being an annual. Climbing plants and species intolerant of low temperatures were less likely to escape. In contrast, the establishment probability was greater if the species belonged to a genus native to Britain and increased as the number of continents in a plant's native range increased. Annual plants had a reduced probability of establishment. Market presence, prices, and the date of introduction are among the socioeconomic factors that have had important effects on the observed course of invasions.
Assuntos
Comércio/estatística & dados numéricos , Demografia , Flores , Desenvolvimento Vegetal , Modelos Logísticos , Dinâmica Populacional , Fatores Socioeconômicos , Especificidade da Espécie , Reino UnidoRESUMO
Protecting national borders against biological invasions is becoming increasingly difficult because those whose actions result in invasions seldom bear legal responsibility for those actions. Invasion costs are often an externality (an unintended side effect) of international trade. Externalities are best dealt with by internalizing them; that is, by getting those who harm society to meet the cost. This is the 'polluter pays principle', which, under current trade rules, is difficult to implement. Tariffs could, however, be used to confront exporters with the costs of their actions, and the right to do this should be embedded in trade agreements. At the same time, international aid could be used to protect donor societies against the inability of some other countries to take appropriate biosecurity measures. The impact of invasions can thus be reduced by tackling their economic externalities.