Regulatory frameworks can facilitate or hinder the potential for genome editing to contribute to sustainable agricultural development.

The advent of new breeding techniques (NBTs), in particular genome editing (GEd), has provided more accurate and precise ways to introduce targeted changes in the genome of both plants and animals. This has resulted in the use of the technology by a wider variety of stakeholders for different applications in comparison to transgenesis. Regulators in different parts of the world are now examining their current frameworks to assess their applicability to these NBTs and their products. We looked at how countries selected from a sample of geographical regions globally are currently handling applications involving GEd organisms and what they foresee as opportunities and potential challenges to acceptance of the technology in their jurisdictions. In addition to regulatory frameworks that create an enabling environment for these NBTs, acceptance of the products by the public is vitally important. We, therefore, suggest that early stakeholder engagement and communication to the public be emphasized to foster public acceptance even before products are ready for market. Furthermore, global cooperation and consensus on issues cutting across regions will be crucial in avoiding regulatory-related bottlenecks that affect global trade and agriculture.

Improving the politics of biotechnological innovations in food security and other sustainable development goals.

The unwarranted interference of some environmental non-governmental organisations (ENGOs) in decision-making over genetically modified (GM) crops has prompted calls for politics to be removed from the regulatory governance of these products. However, regulatory systems are inevitably political because their purpose is to decide whether the use of particular products will help or hinder the delivery of public policy objectives. ENGOs are most able to interfere in regulatory decision-making when policy objectives and decision-making criteria are vague, making the process vulnerable to disruption by organisations that have a distinct agenda. Making regulatory decision-making about GM crops and other green biotechnology more resistant to interference therefore requires better politics not the removal of politics. Better politics begins with political leadership making a case for green biotechnology in achieving food security and other sustainable development goals. Such a policy must involve making political choices and cannot be outsourced to science. Other aspects of better politics include regulatory reform to set policy aims and decision-making criteria that encourage innovation as well as control risk, and engagement with civil society that discusses the values behind attitudes to the application of green biotechnology. In short, green biotechnology for sustainable development needs better politics to counter well-organised opposition, to encourage innovation, and to build the trust of civil society for these policies. Removing politics from regulatory governance would be a gift to ENGOs that are opposed to the use of biotechnology.

Transparency in risk-disproportionate regulation of modern crop-breeding techniques.

Despite over 25 years of safe deployment of genetically engineered crops, the number, complexity, and scope of regulatory studies required for global approvals continue to increase devoid of adequate scientific justification. Recently, there have been calls to further expand the scope of study and data requirements to improve public acceptance. However, increased regulation can actually generate consumer distrust due to the misperception that risks are high. We believe risk-disproportionate regulation as a means to advocate for acceptance of technology is counterproductive, even though some regulatory authorities believe it part of their mandate. To help avoid public distrust, the concept of regulatory transparency to demystify regulatory decision-making should be extended to clearly justifying specific regulatory requirements as: 1) risk-driven (i.e., proportionately addressing increased risk compared with traditional breeding), or 2) advocacy-driven (i.e., primarily addressing consumer concerns and acceptance). Such transparency in the motivation for requiring risk-disproportionate studies would: 1) lessen over-prescriptive regulation, 2) save public and private resources, 3) make beneficial products and technologies available to society sooner, 4) reduce needless animal sacrifice, 5) improve regulatory decision-making regarding safety, and 6) lessen public distrust that is generated by risk-disproportionate regulation.

Problem Formulation for Off-Target Effects of Externally Applied Double-Stranded RNA-Based Products for Pest Control.

Externally applied dsRNA-based biocontrol products may lead to off-target degradation of messenger RNA in target and non-target organisms. For the purposes of regulatory risk assessment of such products, producing a comprehensive catalog of any off-target effects using profiling methods is unnecessary and would be ineffective in supporting decision-making. Instead, problem formulation should derive criteria that indicate acceptable risk and devise a plan to test the hypothesis that the product meets those criteria. The key to effective risk assessment of dsRNA-based biocontrols is determining whether their properties indicate acceptable or unacceptable risk, not whether they arise from on- or off-target effects of dsRNA.

Advancing ecological risk assessment on genetically engineered breeding stacks with combined insect-resistance traits.

To inform the ecological risk assessment (ERA) of a transgenic crop with multiple insecticidal traits combined by conventional breeding (breeding stack), a comparative field study is customarily conducted to compare transgenic protein concentrations in a breeding stack to those in corresponding component single events used in the breeding process. This study tests the hypothesis that transgenic protein expression will not significantly increase due to stacking, such that existing margins of exposure erode to unacceptable levels. Corroboration of this hypothesis allows for the use of existing non-target organism (NTO) effects tests results, where doses were based on the estimated environmental concentrations determined for a component single event. Results from over 20 studies comparing expression profiles of insecticidal proteins produced by commercial events in various combinations of conventionally-bred stacks were examined to evaluate applying previously determined no-observed-effect concentrations (NOECs) to stack ERAs. This paper presents a large number of tests corroborating the hypothesis of no significant increase in insecticidal protein expression due to combination by conventional breeding, and much of the variation in protein expression is likely attributed to genetic and environmental factors. All transgenic protein concentrations were well within conservative margins between exposure and corresponding NOEC. This work supports the conclusion that protein expression data generated for single events and the conservative manner for setting NTO effects test concentrations allows for the transportability of existing NOECs to the ERA of conventionally-bred stacks, and that future tests of the stated hypothesis are no longer critically informative for ERA on breeding stacks.

From Basic Research in Toxicology to a Career in Regulatory Toxicology: An Industry Perspective.

Regulatory toxicology helps define the balance between health risk and societal benefit of chemicals by applying a science-based approach, thus representing a potential career opportunity for scientists involved in biomedical research.

Problem formulation and phenotypic characterisation for the development of novel crops.

Phenotypic characterisation provides important information about novel crops that helps their developers to make technical and commercial decisions. Phenotypic characterisation comprises two activities. Product characterisation checks that the novel crop has the qualities of a viable product-the intended traits have been introduced and work as expected, and no unintended changes have been made that will adversely affect the performance of the final product. Risk assessment evaluates whether the intended and unintended changes are likely to harm human health or the environment. Product characterisation follows the principles of problem formulation, namely that the characteristics required in the final product are defined and criteria to decide whether the novel crop will have these properties are set. The hypothesis that the novel crop meets the criteria are tested during product development. If the hypothesis is corroborated, development continues, and if the hypothesis is falsified, the product is redesigned or its development is halted. Risk assessment should follow the same principles. Criteria that indicate the crop poses unacceptable risk should be set, and the hypothesis that the crop does not possess those properties should be tested. However, risk assessment, particularly when considering unintended changes introduced by new plant breeding methods such as gene editing, often ignores these principles. Instead, phenotypic characterisation seeks to catalogue all unintended changes by profiling methods and then proceeds to work out whether any of the changes are important. This paper argues that profiling is an inefficient and ineffective method of phenotypic characterisation for risk assessment. It discusses reasons why profiling is favoured and corrects some misconceptions about problem formulation.

Using problem formulation to clarify the meaning of weight of evidence and biological relevance in environmental risk assessments for genetically modified crops.

Weight of evidence and biological relevance are important concepts for risk assessment and decision-making over the use of GM crops; however, their meanings are not well defined. We use problem formulation to clarify the definition of these concepts and thereby identify data that are relevant for risk assessment. Problem formulation defines criteria for the acceptability of risk and devises rigorous tests of the hypothesis that the criteria are met. Corroboration or falsification of such hypotheses characterize risk and enable predictable and transparent decisions about whether certain risks from using a particular GM crop are acceptable. Decisions based on a weight of evidence approach use a synthesis of several lines of evidence, whereas a "definitive" approach to risk assessment enables some decisions to be based on the results of a single test. Data are biologically relevant for risk assessment only if they test a hypothesis that is useful for decision-making.

Ecological networks reveal resilience of agro-ecosystems to changes in farming management.

Sustainable management of ecosystems and growth in agricultural productivity is at the heart of the United Nations' Sustainable Development Goals for 2030. New management regimes could revolutionize agricultural production, but require an evaluation of the risks and opportunities. Replacing existing conventional weed management with genetically modified, herbicide-tolerant (GMHT) crops, for example, might reduce herbicide applications and increase crop yields, but remains controversial owing to concerns about potential impacts on biodiversity. Until now, such new regimes have been assessed at the species or assemblage level, whereas higher-level ecological network effects remain largely unconsidered. Here, we conduct a large-scale network analysis of invertebrate communities across 502 UK farm sites to GMHT management in different crop types. We find that network-level properties were overwhelmingly shaped by crop type, whereas network structure and robustness were apparently unaltered by GMHT management. This suggests that taxon-specific effects reported previously did not escalate into higher-level systemic structural change in the wider agricultural ecosystem. Our study highlights current limitations of autecological assessments of effect in agriculture in which species interactions and potential compensatory effects are overlooked. We advocate adopting the more holistic system-level evaluations that we explore here, which complement existing assessments for meeting our future agricultural needs.

A simple problem formulation framework to create the right solution to the right problem.

A systematic approach to formulate consistent, technically robust and scientifically tractable problems will facilitate achieving innovative and effective solutions in risk evaluation. The fundamentals of problem formulation have been adapted from environmental and human health risk assessments. A structured problem formulation enables focus on describing and evaluating the specifics of the problem to be solved, instead of immediately creating solutions. First the problem should be framed to provide clarity and gain agreement on the problem to be addressed, resulting in a specific problem statement. Second the problem is explored in order to transform it into an operational state through questions to answer, hypotheses to test, and represented by a conceptual model. Finally the approach to testing hypotheses is mapped and the analysis plan is developed to address the problem statement. This simple adaptable framework can be applied to any circumstance to resolve a specific problem and describe a path to resolution.

The Integration of Science and Policy in Regulatory Decision-Making: Observations on Scientific Expert Panels Deliberating GM Crops in Centers of Diversity.

Panels of experts with specialized knowledge and experience are often convened to identify and analyze information relevant for risk assessments of GM crops. A perspective on the use of such scientific expert panels is shared here based on panels convened to inform the regulatory strategy for three separate projects developing GM crops for cultivation in Africa: a nutritionally enhanced sorghum, an insect resistant cowpea, and a virus resistant cassava. The panels were convened specifically to consider the risks associated with gene flow from a genetically modified (GM) crop to naturally occurring 'wild' relatives of that crop. In these cases, the experts used problem formulation to identify effects that regulatory authorities may consider to be harmful ("harms") and formulate plausible scenarios that might lead to them, and the availability of information that could determine the likelihood of the steps in the pathway. These panels and the use of problem formulation worked well to gather the existing information and consider the likelihood of harm from gene flow in centers of diversity. However, one important observation from all of these cases is that it is outside the remit of such scientific expert panels to make decisions dependent on policy, such as which harms should be considered and what information should be considered essential in order for a regulatory authority to make a decision about the acceptable level of risk. These experiences of expert panels to inform GM crop risk assessment demonstrate the challenge of integrating science and policy for effective regulatory decision-making.

Policy-Led Comparative Environmental Risk Assessment of Genetically Modified Crops: Testing for Increased Risk Rather Than Profiling Phenotypes Leads to Predictable and Transparent Decision-Making.

We describe two contrasting methods of comparative environmental risk assessment for genetically modified (GM) crops. Both are science-based, in the sense that they use science to help make decisions, but they differ in the relationship between science and policy. Policy-led comparative risk assessment begins by defining what would be regarded as unacceptable changes when the use a particular GM crop replaces an accepted use of another crop. Hypotheses that these changes will not occur are tested using existing or new data, and corroboration or falsification of the hypotheses is used to inform decision-making. Science-led comparative risk assessment, on the other hand, tends to test null hypotheses of no difference between a GM crop and a comparator. The variables that are compared may have little or no relevance to any previously stated policy objective and hence decision-making tends to be ad hoc in response to possibly spurious statistical significance. We argue that policy-led comparative risk assessment is the far more effective method. With this in mind, we caution that phenotypic profiling of GM crops, particularly with omics methods, is potentially detrimental to risk assessment.

When the Whole is Not Greater than the Sum of the Parts: A Critical Review of Laboratory Bioassay Effects Testing for Insecticidal Protein Interactions.

Many studies have been conducted to investigate synergism among insecticidal proteins; however, a consensus on minimal data requirements and interpretation is lacking. While some have concluded that all additive predictive-type models should be abandoned, we advocate that additivity models can remain useful as assessment tools and that an appropriately designed interaction study will never systematically underestimate the existence of synergism, irrespective of which additivity model (or none at all) may be used. To generate the most meaningful synergy assessment datasets in support of safety assessments, we highlight two beneficial steps to follow: (i) select a testing model which is the most consistent with current knowledge regarding the action of the insecticidal proteins and (ii) avoid using bioassay methods which may result in excess response heterogeneity. We also outline other experimental design elements to consider, which may be crucial for conducting future studies of this type. A contrast of underlying testing assumptions associated with the additivity models is provided, along with a comprehensive review of interaction data for Cry1, Cry2, Cry3, Cry9, and Vip3A insecticidal proteins. Our review captures four recurrent findings: i) experiments reporting synergistic interactions are a minority, ii) the degree of synergism reported is low in magnitude, iii) reported interactions are sometimes equivocal/inconclusive due to unconfirmed model assumptions or other bioassay challenges, and iv) due to biological response variation many of the reported interactions may be artefactual. A brief overview of the positioning of interaction testing data in safety assessments of GM food crops is also provided.

Next-Generation Global Biomonitoring: Large-scale, Automated Reconstruction of Ecological Networks.

We foresee a new global-scale, ecological approach to biomonitoring emerging within the next decade that can detect ecosystem change accurately, cheaply, and generically. Next-generation sequencing of DNA sampled from the Earth's environments would provide data for the relative abundance of operational taxonomic units or ecological functions. Machine-learning methods would then be used to reconstruct the ecological networks of interactions implicit in the raw NGS data. Ultimately, we envision the development of autonomous samplers that would sample nucleic acids and upload NGS sequence data to the cloud for network reconstruction. Large numbers of these samplers, in a global array, would allow sensitive automated biomonitoring of the Earth's major ecosystems at high spatial and temporal resolution, revolutionising our understanding of ecosystem change.

A General Approach to Test for Interaction Among Mixtures of Insecticidal Proteins Which Target Different Orders of Insect Pests.

A shift toward transgenic crops which produce combinations of insecticidal proteins has increased the interest (Syngenta Seeds, Inc., Minnetonka, MN) in studying the potential for interactions amongst those proteins. We present a general testing method which accommodates proteins with nonoverlapping spectrums of activity. Our sequential testing approach first investigates groups of the proteins with overlapping activity; e.g., proteins active against Lepidoptera or Coleoptera, respectively. The Colby method is used to test for interactions within each respective group. Subsequently, the mixture of proteins within each group is regarded as a single entity and tests for interactions between the groups (when combined) is conducted using analysis of variance. We illustrate the method using Cry1Ab, Vip3Aa20, and Cry1F (a mixture of proteins active against Lepidoptera), and mCry3A and eCry3.1Ab (a mixture of proteins active against Coleoptera). These insecticidal proteins are produced by Bt11 × MIR162 × TC1507 × MIR604 × 5307 maize. We detected no interactions between Cry1Ab, Vip3Aa20, and Cry1F in tests using larvae of two different lepidopteran species, and possible slight antagonism between mCry3A and eCry3.1Ab with a coleopteran test species. We detected no effect of (eCry3.1Ab + mCry3A) on the potency of (Cry1Ab + Vip3Aa20 + Cry1F) to lepidopteran larvae, and no effect of (Cry1Ab + Vip3Aa20 + Cry1F) on the potency of (mCry3A + eCry3.1Ab) to coleopteran larvae. We discuss implications of these results for characterization of Bt11 × MIR162 × TC1507 × MIR604 × 5307 maize, and the value of the method for characterizing other transgenic crops that produce several insecticidal proteins.

Risk assessment of gene flow from genetically engineered virus resistant cassava to wild relatives in Africa: an expert panel report.

The probability and consequences of gene flow to wild relatives is typically considered in the environmental risk assessment of genetically engineered crops. This is a report from a discussion by a group of experts who used a problem formulation approach to consider existing information for risk assessment of gene flow from cassava (Manihot esculenta) genetically engineered for virus resistance to the 'wild' (naturalized) relative M. glaziovii in East Africa. Two environmental harms were considered in this case: (1) loss of genetic diversity in the germplasm pool, and (2) loss of valued species, ecosystem resources, or crop yield and quality due to weediness or invasiveness of wild relatives. Based on existing information, it was concluded that gene flow will occur, but it is not likely that this will reduce the genetic diversity in the germplasm pool. There is little existing information about the impact of the virus in natural populations that could be used to inform a prediction about whether virus resistance would lead to an increase in reproduction or survival, hence abundance of M. glaziovii. However, an increase in the abundance of M. glaziovii should be manageable, and would not necessarily lead to the identified environmental harms.

[Agricultural biotechnology safety assessment].

Genetically modified (GM) crops were first introduced to farmers in 1995 with the intent to provide better crop yield and meet the increasing demand for food and feed. GM crops have evolved to include a thorough safety evaluation for their use in human food and animal feed. Safety considerations begin at the level of DNA whereby the inserted GM DNA is evaluated for its content, position and stability once placed into the crop genome. The safety of the proteins coded by the inserted DNA and potential effects on the crop are considered, and the purpose is to ensure that the transgenic novel proteins are safe from a toxicity, allergy, and environmental perspective. In addition, the grain that provides the processed food or animal feed is also tested to evaluate its nutritional content and identify unintended effects to the plant composition when warranted. To provide a platform for the safety assessment, the GM crop is compared to non-GM comparators in what is typically referred to as composition equivalence testing. New technologies, such as mass spectrometry and well-designed antibody-based methods, allow better analytical measurements of crop composition, including endogenous allergens. Many of the analytical methods and their intended uses are based on regulatory guidance documents, some of which are outlined in globally recognized documents such as Codex Alimentarius. In certain cases, animal models are recommended by some regulatory agencies in specific countries, but there is typically no hypothesis or justification of their use in testing the safety of GM crops. The quality and standardization of testing methods can be supported, in some cases, by employing good laboratory practices (GLP) and is recognized in China as important to ensure quality data. Although the number of recommended, in some cases, required methods for safety testing are increasing in some regulatory agencies, it should be noted that GM crops registered to date have been shown to be comparable to their nontransgenic counterparts and safe . The crops upon which GM development are based are generally considered safe.