Genetic conversion of a split-drive into a full-drive element

Gene-drive systems offer an important new avenue for spreading beneficial traits into wild populations. Their core components, Cas9 and guide RNA (gRNA), can either be linked within a single cassette (full gene drive, fGD) or provided in two separate elements (split gene drive, sGD) wherein the gRNA-bearing element drives in the presence of an independent static source of Cas9. We previously designed a system engineered to turn split into full gene drives. Here, we provide experimental proof-of-principle for such a convertible system inserted at the spo11 locus, which is recoded to restore gene function. In multigenerational cage studies, the reconstituted spo11 fGD cassette initially drives with slower kinetics than the unlinked sGD element (using the same Mendelian vasa-Cas9 source), but eventually reaches a similar level of final introgression. Different kinetic behaviors may result from transient fitness costs associated with individuals co-inheriting Cas9 and gRNA transgenes during the drive process.

CRISPR gene-drive systems based on Cas9 nickases promote super-Mendelian inheritance in Drosophila

ABSTRACTCRISPR-based gene drive systems can be used to modify entire wild populations due to their ability to bias their own inheritance towards super-Mendelian rates (>100%). Current gene drives contain a Cas9 and a gRNA gene inserted at the location targeted by the gRNA. These gene products are able to cut the opposing wildtype allele, and lead to its replacement with a copy of the gene drive through the homology-directed DNA repair pathway. When this allelic conversion occurs in the germline it leads to the preferential inheritance of the engineered allele — a property that has been proposed to disseminate engineered traits for managing disease-transmitting mosquito populations. Here, we report a novel gene-drive strategy relying on Cas9 nickases which operates by generating staggered paired-nicks in the DNA to promote propagation of the gene drive allele. We show that only when 5’ overhangs are generated, the system efficiently leads to allelic conversion. Further, the nickase gene-drive arrangement produces large stereotyped deletions, providing potential advantages for targeting essential genes. Indeed, the nickase-gene-drive design should expand the options available for gene drive designs aimed at applications in mosquitoes and beyond.

Grundprobleme und Perspektiven des Gentechnikrechts

ZusammenfassungDer vorliegende Beitrag zielt darauf ab, einen Überblick über aktuelle Entwicklungen des Gentechnikrechts zu verschaffen. Neben einigen (immer noch bestehenden) Grundproblematiken im Spannungsfeld zwischen Vorsorgeprinzip und Innovationsbestreben, gibt es sowohl Neuerungen in der europäischen Rechtsprechung, als auch in der technischen und gesellschaftlichen Entwicklung, die vorgestellt und in Kontext gesetzt werden. Einer engeren Untersuchung werden dabei u.a. das Mutagenese-Urteil des EuGH aus dem Jahr 2018 und sogenannte “Gene-Drives” unterzogen.

Propagation of seminal toxins through binary expression gene drives can suppress polyandrous populations

Gene drives can be highly effective in controlling a target population by disrupting a female fertility gene. To spread across a population, these drives require that disrupted alleles be largely recessive so as not to impose too high of a fitness penalty. We argue that this restriction may be relaxed by using a double gene drive design to spread a split binary expression system. One drive carries a dominant lethal/toxic effector alone and the other a transactivator factor, without which the effector will not act. Only after the drives reach sufficiently high frequencies would individuals have the chance to inherit both system components and the effector be expressed. We explore through mathematical modeling the potential of this design to spread dominant lethal/toxic alleles and suppress populations. We show that this system could be implemented to spread engineered seminal proteins designed to kill females, making it highly effective against polyandrous populations.

Philosophy of Technoscience: From Cis-Continental to Trans-Continental

The previous chapters explored how four (interacting and overlapping) continental approaches (dialectics, dialectical materialism, psychoanalysis and phenomenology) offer hints and guidance for coming to terms with the revolutionary dynamics and disruptive impact of contemporary technoscience. Hegelian dialectics provides a conceptual scaffold for developing a comprehensive view of the terrestrial system and even for addressing the Cambrian explosion currently unfolding in laboratories around the globe, as a result of technoscientific developments such as synthetic biology and CRISP-Cas9. Dialectical materialism likewise offers a conceptual framework for addressing the rapidly aggravating disruption of the metabolism between nature and global civilisation, and the ongoing convergence of biosphere and technosphere, exemplified by the synthetic cell. Francophone psychoanalysis, closely aligned with dialectical thinking, adds to our understanding of the specificity of technoscience, both as a practice and as a discourse, where technoscientific research emerges as a questionable vocation driven by a desire to control, but at the same time ostensibly out of control. The dialectical methodology of psychoanalysis was exemplified with the help of case histories, moreover, involving Majorana particles, gene drives, malaria mosquitoes and nude mice. The latter represent technoscientific commodities, exemplifying the assembly-line production of human-made organisms (the commodification of life as such). Subsequently, we demonstrated how Heideggerian phenomenology entails important methodological hints for understanding technoscientific artefacts against the backdrop of technoscience as a mobilising force and as a global enterprise. And finally, we outlined how Teilhard’s views on the genesis of consciousness, self-consciousness and hyperconsciousness retrieve the historical (dialectical) dimension of phenomenology, thus allowing us to assess the present as a global unfolding of the noosphere.

New developments in the field of genomic technologies and their relevance to conservation management

Recent technological advances in the field of genomics offer conservation managers and practitioners new tools to explore for conservation applications. Many of these tools are well developed and used by other life science fields, while others are still in development. Considering these technological possibilities, choosing the right tool(s) from the toolbox is crucial and can pose a challenging task. With this in mind, we strive to inspire, inform and illuminate managers and practitioners on how conservation efforts can benefit from the current genomic and biotechnological revolution. With inspirational case studies we show how new technologies can help resolve some of the main conservation challenges, while also informing how implementable the different technologies are. We here focus specifically on small population management, highlight the potential for genetic rescue, and discuss the opportunities in the field of gene editing to help with adaptation to changing environments. In addition, we delineate potential applications of gene drives for controlling invasive species. We illuminate that the genomic toolbox offers added benefit to conservation efforts, but also comes with limitations for the use of these novel emerging techniques.

Modeling the efficacy of CRISPR gene drive for schistosomiasis control

CRISPR gene drives could revolutionize the control of infectious diseases by accelerating the spread of engineered traits that limit parasite transmission in wild populations. While much effort has been spent developing gene drives in mosquitoes, gene drive technology in molluscs has received little attention despite the role of freshwater snails as obligate, intermediate hosts of parasitic flukes causing schistosomiasis -- a disease of poverty affecting more than 200 million people worldwide. A successful drive in snails must overcome self-fertilization, which prevents a drive's spread. Simultaneous hermaphroditism is a feature of snails -- distinct from gene drive model organisms -- and is not yet incorporated in gene drive models of disease control. Here we developed a novel population genetic model accounting for snails' sexual and asexual reproduction, susceptibility to parasite infection regulated by multiple alleles, fitness differences between genotypes, and a range of drive characteristics. We then integrated this model with an epidemiological model of schistosomiasis transmission and snail population dynamics. Simulations showed that gene drive establishment can be hindered by a variety of biological and ecological factors, including selfing. However, our model suggests that, under a range of conditions, gene drive mediated immunity in snails could maintain rapid disease reduction achieved by annual chemotherapy treatment of the human population, leading to long-term elimination. These results indicate that gene drives, in coordination with existing public health measures, may become a useful tool to reduce schistosomiasis burden in selected transmission settings with effective CRISPR construct design and close evaluation of the genetic and ecological landscape.

Modeling homing suppression gene drive in haplodiploid organisms

Gene drives have shown great promise for suppression of pest populations. These engineered alleles can function by a variety of mechanisms, but the most common is the CRISPR homing drive, which converts wild-type alleles to drive alleles in the germline of heterozygotes. Some potential target species are haplodiploid, in which males develop from unfertilized eggs and thus have only one copy of each chromosome. This prevents drive conversion, a substantial disadvantage compared to diploids where drive conversion can take place in both sexes. Here, we study the characteristics of homing suppression gene drives in haplodiploids and find that a drive targeting a female fertility gene could still be successful. However, such drives are less powerful than in diploids. They are substantially more vulnerable to high resistance allele formation in the embryo due to maternally deposited Cas9 and gRNA and also to somatic cleavage activity. Examining models of continuous space where organisms move over a landscape, we find that haplodiploid suppression drives surprisingly perform nearly as well as in diploids, possibly due to their ability to spread further before inducing strong suppression. Together, these results indicate that gene drive can potentially be used to effectively suppress haplodiploid populations.