Publications by authors named "Joachim De Jonghe"

Understanding the rapidly evolving landscape of single-cell and spatial omic technologies is crucial for advancing biomedical research and drug development. We provide a living review of both mature and emerging commercial platforms, highlighting key methodologies and trends shaping the field. This review spans from foundational single-cell technologies such as microfluidics and plate-based methods to newer approaches like combinatorial indexing; on the spatial side, we consider next-generation sequencing and imaging-based spatial transcriptomics.

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  • Precision medicine relies on understanding genetic variants that cause disease and their effects, exemplified by the von Hippel-Lindau (VHL) gene involved in tumor suppression.
  • VHL mutations are linked to specific types of tumors, like clear cell renal cell carcinoma, necessitating refined methods to assess their consequences.
  • Researchers developed a technique to analyze nearly all single-nucleotide variants in VHL, leading to the identification of key pathogenic variants and enhancing the ability to classify these genetic changes in clinical settings.
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  • Droplet microfluidic methods have improved the efficiency of single-cell sequencing but face challenges like increased background noise and lower RNA capture rates due to the lack of effective cell enrichment strategies.
  • The presented methodology uses fluorescence-activated droplet sorting to isolate droplets containing viable or specific cell types and employs picoinjection for multi-step processes, enhancing gene detection by five times and reducing noise by up to 50%.
  • This approach successfully creates a high-quality molecular atlas of mouse brain development and nascent RNA transcription during organogenesis, and can be adapted for various other droplet-based workflows to achieve cost-effective and precise single-cell profiling.
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  • Biomechanical cues are crucial for embryonic development and cell differentiation, and studying these can reveal how physical stimuli influence gene expression during early mammalian development.
  • By using microfluidic techniques to encapsulate mouse embryonic stem cells, researchers found that Plakoglobin (Jup), a key protein, enhances the network responsible for maintaining naive pluripotency.
  • The study highlights Plakoglobin's role as a mechanosensitive regulator, suggesting that its expression during blastocyst formation in both human and mouse embryos is vital for understanding cell fate transitions influenced by the physical environment.
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Single-cell RNA sequencing (scRNA-seq) is a powerful technique for describing cell states. Identifying the spatial arrangement of these states in tissues remains challenging, with the existing methods requiring niche methodologies and expertise. Here, we describe segmentation by exogenous perfusion (SEEP), a rapid and integrated method to link surface proximity and environment accessibility to transcriptional identity within three-dimensional (3D) disease models.

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Mammalian embryos sequentially differentiate into trophectoderm and an inner cell mass, the latter of which differentiates into primitive endoderm and epiblast. Trophoblast stem (TS), extraembryonic endoderm (XEN) and embryonic stem (ES) cells derived from these three lineages can self-assemble into synthetic embryos, but the mechanisms remain unknown. Here, we show that a stem cell-specific cadherin code drives synthetic embryogenesis.

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Embryonic stem (ES) cells can undergo many aspects of mammalian embryogenesis in vitro, but their developmental potential is substantially extended by interactions with extraembryonic stem cells, including trophoblast stem (TS) cells, extraembryonic endoderm stem (XEN) cells and inducible XEN (iXEN) cells. Here we assembled stem cell-derived embryos in vitro from mouse ES cells, TS cells and iXEN cells and showed that they recapitulate the development of whole natural mouse embryo in utero up to day 8.5 post-fertilization.

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  • Traditional single-cell transcriptome sequencing only captures a limited portion of transcripts by focusing on polyadenylated RNA, missing many important non-coding and non-polyadenylated transcripts.
  • The new VASA-seq method allows for the analysis of the entire transcriptome in single cells by fragmenting and tailing all RNA after cell lysis, and it works with both plate-based and droplet microfluidic systems.
  • Applying VASA-seq to over 30,000 single cells in developing mouse embryos revealed key findings such as novel cell type markers, insights into blood maturation trajectories, and extensive alternative splicing during development.
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The development of mouse embryos can be partially recapitulated by combining embryonic stem cells (ESCs), trophoblast stem cells (TS), and extra-embryonic endoderm (XEN) stem cells to generate embryo-like structures called ETX embryos. Although ETX embryos transcriptionally capture the mouse gastrula, their ability to recapitulate complex morphogenic events such as gastrulation is limited, possibly due to the limited potential of XEN cells. To address this, we generated ESCs transiently expressing transcription factor Gata4, which drives the extra-embryonic endoderm fate, and combined them with ESCs and TS cells to generate induced ETX embryos (iETX embryos).

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Mammalian blastocysts comprise three distinct cell lineages essential for development beyond implantation: the pluripotent epiblast, which generates the future embryo, and surrounding it the extra-embryonic primitive endoderm and the trophectoderm tissues. Embryonic stem cells can reintegrate into embryogenesis but contribute primarily to epiblast lineages. Here, we show that mouse embryonic stem cells cultured under extended pluripotent conditions (EPSCs) can be partnered with trophoblast stem cells to self-organize into blastocyst-like structures with all three embryonic and extra-embryonic lineages.

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Low-cost shotgun DNA sequencing is transforming the microbial sciences. Sequencing instruments are so effective that sample preparation is now the key limiting factor. Here, we introduce a microfluidic sample preparation platform that integrates the key steps in cells to sequence library sample preparation for up to 96 samples and reduces DNA input requirements 100-fold while maintaining or improving data quality.

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Geobacillus thermoglucosidasius is a Gram-positive thermophile of industrial interest that exhibits rapid growth and can utilize a variety of plant-derived feedstocks. It is an attractive chassis organism for high temperature biotechnology and synthetic biology applications but is currently limited by a lack of available genetic tools. Here we describe a set of modular shuttle vectors, including a promoter library and reporter proteins.

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Overlap-directed DNA assembly methods allow multiple DNA parts to be assembled together in one reaction. These methods, which rely on sequence homology between the ends of DNA parts, have become widely adopted in synthetic biology, despite being incompatible with a key principle of engineering: modularity. To answer this, we present MODAL: a Modular Overlap-Directed Assembly with Linkers strategy that brings modularity to overlap-directed methods, allowing assembly of an initial set of DNA parts into a variety of arrangements in one-pot reactions.

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