Targeted mutagenesis in a human-parasitic nematode.

Parasitic nematodes infect over 1 billion people worldwide and cause some of the most common neglected tropical diseases. Despite their prevalence, our understanding of the biology of parasitic nematodes has been limited by the lack of tools for genetic intervention. In particular, it has not yet been possible to generate targeted gene disruptions and mutant phenotypes in any parasitic nematode. Here, we report the development of a method for introducing CRISPR-Cas9-mediated gene disruptions in the human-parasitic threadworm Strongyloides stercoralis. We disrupted the S. stercoralis twitchin gene unc-22, resulting in nematodes with severe motility defects. Ss-unc-22 mutations were resolved by homology-directed repair when a repair template was provided. Omission of a repair template resulted in deletions at the target locus. Ss-unc-22 mutations were heritable; we passed Ss-unc-22 mutants through a host and successfully recovered mutant progeny. Using a similar approach, we also disrupted the unc-22 gene of the rat-parasitic nematode Strongyloides ratti. Our results demonstrate the applicability of CRISPR-Cas9 to parasitic nematodes, and thereby enable future studies of gene function in these medically relevant but previously genetically intractable parasites

Matterport3D: Learning from RGB-D Data in Indoor Environments

Access to large, diverse RGB-D datasets is critical for training RGB-D scene understanding algorithms. However, existing datasets still cover only a limited number of views or a restricted scale of spaces. In this paper, we introduce Matterport3D, a large-scale RGB-D dataset containing 10,800 panoramic views from 194,400 RGB-D images of 90 building-scale scenes. Annotations are provided with surface reconstructions, camera poses, and 2D and 3D semantic segmentations. The precise global alignment and comprehensive, diverse panoramic set of views over entire buildings enable a variety of supervised and self-supervised computer vision tasks, including keypoint matching, view overlap prediction, normal prediction from color, semantic segmentation, and region classification

Simultaneous precise editing of multiple genes in human cells

Abstract. When double-strand breaks are introduced in a genome by CRISPR they are repaired either by non-homologous end joining (NHEJ), which often results i

Poverty and development : the Human Development Report and the World Development Report, 1990

After the"adjustment decade"of the 1980s, attention in the 1990s seems to be turning once again to longer-term issues of development - particularly of poverty alleviation. Just as the 1980s were heralded by a series of reports on adjustment, so the 1990s have seen two major reports on poverty: World Development Report 1990: Poverty, by the World Bank, and Human Development Report 1990 by the UNDP. The author presents an overview of conceptual issues and the best policies for alleviating poverty, based on a review of these two reports. He poses basic questions on the definition and measurement of poverty, looks at what has actually happened to poverty in developing countries in the last three decades, and reviews policies to help alleviate poverty. The consensus represented in these two reports, he concludes, offers hope that the polarization of policy analysts into"camps"is a thing of the past - and that policies for the 1990s can be built on fundamental agreement about the basics: (a) that poverty alleviation requires growth, but growth is not enough; (b) that growth must be broad-based and labor-intensive, and must go hand in hand with targeted basic social expenditures; and (c) that the international community must do its share, by supporting these efforts in the 1990s through greatly increased capital flows to development countries.Poverty Assessment,Health Economics&Finance,Environmental Economics&Policies,Achieving Shared Growth,Governance Indicators

Measuring well-being with social indicators, HDIs, PQLI, and BWI for 133 countries for 1975, 1980, 1985, 1988, and 1992

Well Being;Measurement;Social Indicators;welfare economics

Quantifying protein densities on cell membranes using super-resolution optical fluctuation imaging

Surface molecules, distributed in diverse patterns and clusters on cell membranes, influence vital functions of living cells. It is therefore important to understand their molecular surface organisation under different physiological and pathological conditions. Here, we present a model-free, quantitative method to determine the distribution of cell surface molecules based on TIRF illumination and super-resolution optical fluctuation imaging (SOFI). This SOFI-based approach is robust towards single emitter multiple-blinking events, high labelling densities and high blinking rates. In SOFI, the molecular density is not based on counting events, but results as an intrinsic property due to the correlation of the intensity fluctuations. The effectiveness and robustness of the method was validated using simulated data, as well as experimental data investigating the impact of palmitoylation on CD4 protein nanoscale distribution in the plasma membrane of resting T cells.Comment: 9 pages, 3 figures plus Supplementary Informatio

Extension of the crRNA enhances Cpf1 gene editing in vitro and in vivo.

Engineering of the Cpf1 crRNA has the potential to enhance its gene editing efficiency and non-viral delivery to cells. Here, we demonstrate that extending the length of its crRNA at the 5 end can enhance the gene editing efficiency of Cpf1 both in cells and in vivo. Extending the 5 end of the crRNA enhances the gene editing efficiency of the Cpf1 RNP to induce non-homologous end-joining and homology-directed repair using electroporation in cells. Additionally, chemical modifications on the extended 5 end of the crRNA result in enhanced serum stability. Also, extending the 5 end of the crRNA by 59 nucleotides increases the delivery efficiency of Cpf1 RNP in cells and in vivo cationic delivery vehicles including polymer nanoparticle. Thus, 5 extension and chemical modification of the Cpf1 crRNA is an effective method for enhancing the gene editing efficiency of Cpf1 and its delivery in vivo

Accountability in Global Governance

human development, democracy

National Adaptation Strategies to Climate Change Impacts: A Case Study of Mozambique

human development, climate change