We report a robust, versatile approach called CRISPR live-cell fluorescent in situ hybridization (LiveFISH) using fluorescent oligonucleotides for genome tracking in a broad range of cell types, including primary cells. An intrinsic stability switch of CRISPR guide RNAs enables LiveFISH to accurately detect chromosomal disorders such as Patau syndrome in prenatal amniotic fluid cells and track multiple loci in human T lymphocytes. In addition, LiveFISH tracks the real-time movement of DNA double-strand breaks induced by CRISPR-Cas9–mediated editing and consequent chromosome translocations. Finally, by combining Cas9 and Cas13 systems, LiveFISH allows for simultaneous visualization of genomic DNA and RNA transcripts in living cells. The LiveFISH approach enables real-time live imaging of DNA and RNA during genome editing, transcription, and rearrangements in single cells.

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Visualizing the real-time dynamics of genome rearrangement in single living cells is core to studying genomics and diagnostics. Here, we report a robust, versatile approach named CRISPR Live-cell fluorescent in situ hybridization (LiveFISH) for multi-locus genome tracking and cytogenetic detection in a broad variety of cell types including primary cells. LiveFISH utilizes an intrinsic stability switch of CRISPR guide RNAs, which enables efficient and accurate detection of chromosomal disorders such as Patau Syndrome in prenatal amniotic fluid cells and allows multi-locus tracking in human T lymphocytes. Using LiveFISH, we are able to detect and track real-time spatiotemporal dynamics of non-homologous endogenous chromosome translocations induced by gene editing. This new approach enables FISH imaging in living primary cells, which can provide useful insights into the spatiotemporal changes of genome organization and rearrangements in normal and diseased primary cells and will enable fast cytogenetic visualization of various gene-editing associated chromosomal translocations. Cell Sciences Imaging Facility for microscope usage, CHEM-H Macromolecular Structure Knowledge Center for protein purification facilities, and Coriell Cell Repositories for providing cell resources.

Human-driven changes on this planet have been giving rise to global warming, social instability, civil wars, and acts of terrorism. The existing system of global governance is not equipped to effectively address these enormous challenges. It is slow where one must move quickly, favors bureaucracy and politics over authentic deliberations and effective interventions, and caters to power-brokers and mega-corporations. The world therefore needs a model of global governance that serves to make and implement collectively binding decisions that acknowledge the interests of all those affected, including future generations. This governance model must coordinate the work of great (e.g. national) powers, and at the same time enable billions of people to bring their intelligence and creativity to bear on these challenges. In many ways, the quest for a new system of global governance is a grand societal challenge in itself. In this paper, we draw on idealized design to develop an ideal model of global governance and explore the collective search and experimentation efforts it implies. This so-called United People (UP) model involves a circular hierarchy in which power and communication flow in ways that help the global community to effectively address transnational challenges and problems. It involves several, relatively small, governance bodies—rather than a large parliamentary assembly that tends to cripple responsive decision-making. The UP model also serves to effectively uncover and address power abuse, simplify the financial household of global governance, and support systemic forms of collaboration with NGOs and other organizations.