Genetic engineering has a wide range of cultural, economic, and ethical implications, yet it has become almost an article of faith that regulatory decisions about biotechnology be based only on ...evidence of specific quantifiable risks; to consider anything else is said to "politicize" regulation. In this study of social protest against genetically engineered food, Abby Kinchy turns the conventional argument on its head. Rather than consider politicization of the regulatory system, she takes a close look at the scientization of public debate about the "contamination" of crops resulting from pollen drift and seed mixing. Advocates of alternative agriculture confront the scientization of this debate by calling on international experts, carrying out their own research, questioning regulatory science in court, building alternative markets, and demanding that their governments consider the social and economic impacts of the new technologies. Kinchy focuses on social conflicts over canola in Canada and maize in Mexico, drawing out their linkages to the global food system and international environmental governance. The book ultimately demonstrates the shortcomings of dominant models of scientific risk governance, which marginalize alternative visions of rural livelihoods and sustainable food production.The hardcover edition does not include a dust jacket.
Keywords: alpha-synuclein; nonmotor symptoms; psychosis; sex differences; dopamine Background Parkinson's disease psychosis is a prevalent yet underreported and understudied nonmotor manifestation of ...Parkinson's disease and, arguably, the most debilitating. It is unknown if alpha-synuclein plays a role in psychosis, and if so, this endophenotype may be crucial for elucidating the neurodegenerative process. Objectives We sought to dissect the underlying neurobiology of novelty-induced hyperactivity, reminiscent of psychosis-like behavior, in human alpha-synuclein BAC rats. Results Herein, we demonstrate a prodromal psychosis-like phenotype, including late-onset sensorimotor gating disruption, striatal hyperdopaminergic signaling, and persistent novelty-induced hyperactivity (up to 18months), albeit reduced baseline locomotor activity, that is augmented by d-amphetamine and reversed by classical and atypical antipsychotics. MicroRNA-mediated alpha-synuclein downregulation in the ventral midbrain rescues the hyperactive phenotype and restores striatal dopamine levels. This phenotype is accompanied by an abundance of age-, brain region- and gene dose-dependent aberrant alpha-synuclein, including hyperphosphorylation, C-terminal truncation, aggregation pathology, and mild nigral neurodegeneration (27%). Conclusions Our findings demonstrate a potential role of alpha-synuclein in Parkinson's disease psychosis and provide evidence of region-specific perturbations prior to neurodegeneration phenoconversion. The reported phenotype coincides with the latest clinical findings that suggest a premotor hyperdopaminergic state may occur, while at the same time, premotor psychotic symptoms are increasingly being recognized. c 2020 International Parkinson and Movement Disorder Society Article Note: Relevant conflict of interest/financial disclosure: The authors report no conflict of interest. Funding agencies: This project was funded by an FP7-HEALTH MULTISYN (602646) awarded to L.S. and a grant awarded to A.P. from the General Secretariat for Research and Technology (GSRT) and the Hellenic Foundation for Research and Innovation (HFRI) (Code: 1855). CAPTION(S): FIG. S1. Quantification of human monomeric and phosphorylated alpha-synuclein (AS) in AS BAC rats in the hippocampus, frontal cortex, striatum, and ventral midbrain. Quantification of Western immunoblots of Triton-X (TX)- (left) and SDS-soluble (right) fractions (blots depicted in Fig. 1). Expression of human monomeric AS (4B12) and phosphorylated AS (at serine 129), respectively, in the hippocampus (A, B), frontal cortex (C, D), striatum (E, F), and midbrain (G, H) at 3, 6, 9, and 12months; AS levels normalized to gamma-tubulin levels and to the positive control "rest brain" sample. (I) Representative immunofluorescence images of phosphorylated AS at Ser129 (pAS Ser129, green, in TH+ nigral neurons, red, in 10-month-old WT (top) and AS-BAC (bottom 2 panels) rats; merged images include DAPI (blue) nuclear staining. Scale bar: 25MUm. Comparison of BAC/WT fold-difference of total monomeric AS levels (human and endogenous AS-C20 antibody) in the TX-soluble (J) and SDS-soluble (K) fraction at 3-12 months in all brain regions (n = 3/group. A-H: One-way ANOVA; I, J: 2-way ANOVA. *P<0.05, brain region comparison. FIG. S2. Monoamine tissue content in the cortex, hippocampus, and amygdala. Noradrenaline (NA), dopamine (DA), and serotonin (5HT) levels (ng/mg wet tissue weight) did not differ between alpha-synuclein (AS) BAC and wild-type (WT) rats at all ages in the cortex (A), hippocampus (B), and amygdala (C). A-C: Two-way ANOVA, n = 3-7/group. FIG. S3. Tyrosine hydroxylase immunoblotting. Striatal tyrosine hydroxylase (TH) levels remained unchanged in alpha-synuclein (AS) BAC rats when measured with Western immunoblotting, as depicted with representative blots (left) and quantification (right) in the (A) TX-soluble and (B) SDS-soluble fraction (unpaired Student t test). FIG. S4. Open-field horizontal activity in 15-minute intervals. Female (left) and male (right) human alpha-synuclein (AS) BAC rats exhibited enhanced horizontal locomotor activity (distance traveled cm) in a novel open field over a 60-minute period, expressed in 15-minute intervals at 3 months (A), 6 months (B), 12 months (C), and 18 months (D) compared with wild-type (WT) rats. Decreased locomotor activity over time signifies a pattern of habituation similar to WT rats. Overall, male AS BAC rats exhibited a more robust behavioral profile. A-D: Three-way ANOVA; *P<0.05; **P<0.01; ***P<0.001. FIG. S5. Open-field vertical activity in 15-minute intervals. Female (left) and male (right) human alpha-synuclein (AS) BAC rats exhibit enhanced vertical locomotor activity (rearing frequency) in a novel open field over a 60-minute period, expressed in 15-minute intervals at 3 months (A), 6 months (B), 12 months (C), and 18 months (D) compared with wild-type (WT) rats. Decreased locomotor activity over time signifies a similar pattern of habituation as WT rats. Overall, male AS BAC rats exhibited a more robust behavioral profile. A-D: Three-way ANOVA; *P<0.05; **P<0.01; ***P<0.001. FIG. S6. Prepulse inhibition (PPI) with a prepulse of 75, 80, 85, and 90dB in female (left) and male (right) human alpha-synuclein (AS) BAC and wild-type (WT) rats at 3 months (A), 6 months (B), 9 months (C), and 12 months (D). Male AS BAC rats exhibited a sound intensity-dependent (85-dB prepulse) PPI deficit (% PPI) at 12 months (average PPI for all prepulse-pulse pairings depicted in Fig. 4I). A-D: Repeated-measures ANOVA; **P<0.01. FIG. S7. Pimavanserin's effects on locomotor activity. Locomotor activity in an open field is expressed as horizontal activity (distance traveled cm) in 3-month male WT and AS BAC rats following pimavanserin administration at 0.3mg/kg subcutaneously (A) or 1mg/kg, subcutaneously (B). Pimavanserin does not appear to significantly modify either AS BAC or WT rat locomotor activity. A, B: Two-way ANOVA. FIG. S8. Targeting of virus injections in the ventral midbrain. (A) GFP-injected (top) and GFP-tagged human alpha-synuclein miRNA-injected (miR AS-GFP) human alpha-synuclein (AS BAC) in the ventral tegmental area (VTA) leading to viral expression (GFP-green) throughout the VTA in tyrosine hydroxylase-stained (TH+) neurons (red). The miR AS led to decreased human AS (pseudocolor) expression in the VTA. (B) Representative images showing the targeting of GFP-tagged (green) viral injections in AS BAC rats in the ventral tegmental area (VTA) that led to expression of virus mainly in TH+ neurons (red) in the VTA but also in the substantia nigra (top) and expression in TH+ afferents in both the dorsal and ventral striata (bottom). DAPI nuclear staining is depicted in the first column, GFP in the second column, TH neuron/afferent staining in the third column, human AS (211 antibody) in the fourth column (A only), and merged images in the fourth (B) or fifth (A) column. A, B: top in B taken with 10x objective lens and bottom in B with a tile scan. FIG. S9. Genotype-phenotype colony comparison. Quantitative gene expression analysis of human alpha-synuclein (alpha-syn) between originating laboratory (Germany, denoted as DE) and our labpratpru (Greece, denoted as GR) showing 25% greater copy number in DE BAC rats (A). Brain tissue of BAC rats or wild-type (WT) controls was lysed, and 2 protein fractions were prepared - TX-soluble fraction (striatum B, midbrain D) and SDS-soluble fraction (striatum C, midbrain E). Membranes were probed for 3 different alpha-syn antibodies (pS129, 4B12 human, and Syn1 total) or tyrosine hydroxylase (TH) - representative blots on the left, quantification on the right. Relative density for monomeric alpha-syn (15kDa) was normalized to the intensity of the gamma-tubulin band (40kDa). All forms of alpha-syn were greater in DE BAC rats in the striatum in both fractions, whereas in the midbrain, pS129 alpha-syn levels were increased in the SDS-soluble fraction and TH levels greatly reduced in DE BAC rats, indicative of neurodegeneration. A-E: Unpaired Student t test, *P<0.05; **P<0.01, ***P<0.001. Appendix S1. Supporting information Byline: Alexia Polissidis, Maria Koronaiou, Vasia Kollia, Effrosyni Koronaiou, Modestos Nakos-Bimpos, Marios Bogiongko, Sofia Vrettou, Katerina Karali, Nicolas Casadei, Olaf Riess, Sergio P. Sardi, Maria Xilouri, Leonidas Stefanis
But although CRISPR has much to offer, some scientists are worried that the field's breakneck pace leaves little time for addressing the ethical and safety concerns such experiments can raise. The ...problem was thrust into the spotlight in April, when news broke that scientists had used CRISPR to engineer human embryos (see Nature 520, 593-595; 2015).
The diversity, modularity, and efficacy of CRISPR-Cas systems are driving a biotechnological revolution. RNA-guided Cas enzymes have been adopted as tools to manipulate the genomes of cultured cells, ...animals, and plants, accelerating the pace of fundamental research and enabling clinical and agricultural breakthroughs. We describe the basic mechanisms that set the CRISPR-Cas toolkit apart from other programmable gene-editing technologies, highlighting the diverse and naturally evolved systems now functionalized as biotechnologies. We discuss the rapidly evolving landscape of CRISPR-Cas applications, from gene editing to transcriptional regulation, imaging, and diagnostics. Continuing functional dissection and an expanding landscape of applications position CRISPR-Cas tools at the cutting edge of nucleic acid manipulation that is rewriting biology.
Flowering is an essential part of the productive process, and flowering time is determined by endogenous genetic components and many ambient factors. SHORT VEGETATIVE PHASE (SVP), a MADS-box ...transcription factor, regulates floral transition by repressing floral integrator genes and is involved in ABA-mediated drought stress. In this study, we transformed the poplar (Populus) clone “84K” with the SVP-Like gene, while stable overexpression transgenic lines were obtained. Transcriptome analysis of the leaves of the transgenic lines and WT (Wide Type) poplars revealed that a total of 477 genes showed significantly altered expression, overexpressing SVL genes, including 342 upregulated and 135 downregulated genes. Ten subclusters in DEGs were analyzed, and KEGG terms of the largest subcluster were associated with two key pathways: hormone-related genes and glutathione metabolism. Meanwhile, many transcriptional factors were involved. Our results are helpful for in-depth analysis of the MADS transcriptional factor in poplars. This work provides the basis for studying woody plant growth, and development and molecular mechanisms responded to environmental stresses.
Targeted modification of the pig genome can be challenging. Recent applications of the CRISPR/Cas9 system hold promise for improving the efficacy of genome editing. When a designed CRISPR/Cas9 system ...targeting CD163 or CD1D was introduced into somatic cells, it was highly efficient in inducing mutations. When these mutated cells were used with somatic cell nuclear transfer, offspring with these modifications were created. When the CRISPR/Cas9 system was delivered into in vitro produced presumptive porcine zygotes, the system was effective in creating mutations in eGFP, CD163, and CD1D (100% targeting efficiency in blastocyst stage embryos); however, it also presented some embryo toxicity. We could also induce deletions in CD163 or CD1D by introducing two types of CRISPRs with Cas9. The system could also disrupt two genes, CD163 and eGFP, simultaneously when two CRISPRs targeting two genes with Cas9 were delivered into zygotes. Direct injection of CRISPR/Cas9 targeting CD163 or CD1D into zygotes resulted in piglets that have mutations on both alleles with only one CD1D pig having a mosaic genotype. We show here that the CRISPR/Cas9 system can be used by two methods. The system can be used to modify somatic cells followed by somatic cell nuclear transfer. System components can also be used in in vitro produced zygotes to generate pigs with specific genetic modifications.
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