Search

Your search keyword '"Jens Puschhof"' showing total 4 results
Start Over Author "Jens Puschhof" Journal nature communications
4 results on '"Jens Puschhof"'

1. Development of Plasmodium falciparum liver-stages in hepatocytes derived from human fetal liver organoid cultures

Author

Annie S. P. Yang, Devanjali Dutta, Kai Kretzschmar, Delilah Hendriks, Jens Puschhof, Huili Hu, Kim E. Boonekamp, Youri van Waardenburg, Susana M. Chuva de Sousa Lopes, Geert-Jan van Gemert, Johannes H. W. de Wilt, Teun Bousema, Hans Clevers, and Robert W. Sauerwein

Subjects
Science
Abstract

Abstract Plasmodium falciparum (Pf) parasite development in liver represents the initial step of the life-cycle in the human host after a Pf-infected mosquito bite. While an attractive stage for life-cycle interruption, understanding of parasite-hepatocyte interaction is inadequate due to limitations of existing in vitro models. We explore the suitability of hepatocyte organoids (HepOrgs) for Pf-development and show that these cells permitted parasite invasion, differentiation and maturation of different Pf strains. Single-cell messenger RNA sequencing (scRNAseq) of Pf-infected HepOrg cells has identified 80 Pf-transcripts upregulated on day 5 post-infection. Transcriptional profile changes are found involving distinct metabolic pathways in hepatocytes with Scavenger Receptor B1 (SR-B1) transcripts highly upregulated. A novel functional involvement in schizont maturation is confirmed in fresh primary hepatocytes. Thus, HepOrgs provide a strong foundation for a versatile in vitro model for Pf liver-stages accommodating basic biological studies and accelerated clinical development of novel tools for malaria control.

Published
2023
Full Text
View/download PDF

2. A turquoise fluorescence lifetime-based biosensor for quantitative imaging of intracellular calcium

Author

Franka H. van der Linden, Eike K. Mahlandt, Janine J. G. Arts, Joep Beumer, Jens Puschhof, Saskia M. A. de Man, Anna O. Chertkova, Bas Ponsioen, Hans Clevers, Jaap D. van Buul, Marten Postma, Theodorus W. J. Gadella, and Joachim Goedhart

Subjects
Science
Abstract

Currently, genetically encoded calcium indicators are not suitable for direct quantification. Here the authors engineer a fluorescence lifetime imaging calcium biosensor, Turquoise Calcium Fluorescence LIfeTime Sensor (Tq-Ca-FLITS), and measure intracellular calcium concentrations in human-derived organoids.

Published
2021
Full Text
View/download PDF

3. A CRISPR/Cas9 genetically engineered organoid biobank reveals essential host factors for coronaviruses

Author

Joep Beumer, Maarten H. Geurts, Mart M. Lamers, Jens Puschhof, Jingshu Zhang, Jelte van der Vaart, Anna Z. Mykytyn, Tim I. Breugem, Samra Riesebosch, Debby Schipper, Petra B. van den Doel, Wim de Lau, Cayetano Pleguezuelos-Manzano, Georg Busslinger, Bart L. Haagmans, and Hans Clevers

Subjects
Science
Abstract

Rapid identification of host genes essential for virus replication may expedite the generation of therapeutic interventions. Here the authors generate mutant clonal intestinal organoids for 19 host genes previously implicated in coronavirus biology and identify the cell surface protease TMPRSS2 as a potential therapeutic target.

Published
2021
Full Text
View/download PDF

4. A CRISPR/Cas9 genetically engineered organoid biobank reveals essential host factors for coronaviruses

Author

Jelte van der Vaart, Bart L. Haagmans, Anna Z Mykytyn, Georg A. Busslinger, Tim I Breugem, Mart M. Lamers, Maarten H. Geurts, Wim de Lau, Cayetano Pleguezuelos-Manzano, Joep Beumer, Jingshu Zhang, Debby Schipper, Petra B. van den Doel, Hans Clevers, Jens Puschhof, Samra Riesebosch, Virology, and Hubrecht Institute for Developmental Biology and Stem Cell Research

Subjects
CRISPR-Cas9 genome editing, Proteases, Middle East respiratory syndrome coronavirus, Science, Dipeptidyl Peptidase 4, viruses, Mutant, General Physics and Astronomy, Serine Endopeptidases/genetics, Computational biology, Biology, medicine.disease_cause, Virus Replication, General Biochemistry, Genetics and Molecular Biology, Article, Stem-cell biotechnology, Cell Line, Dipeptidyl Peptidase 4/genetics, SDG 3 - Good Health and Well-being, medicine, Organoid, Organoids/metabolism, CRISPR, Humans, Gene, Coronavirus, Biological Specimen Banks, Multidisciplinary, SARS-CoV-2, Intestinal stem cells, Serine Endopeptidases, virus diseases, COVID-19, General Chemistry, Organoids, Angiotensin-Converting Enzyme 2/genetics, Viral replication, Genetic engineering, Middle East Respiratory Syndrome Coronavirus, Angiotensin-Converting Enzyme 2, CRISPR-Cas Systems, Transcriptome, Genetic screen
Abstract

Rapid identification of host genes essential for virus replication may expedite the generation of therapeutic interventions. Genetic screens are often performed in transformed cell lines that poorly represent viral target cells in vivo, leading to discoveries that may not be translated to the clinic. Intestinal organoids are increasingly used to model human disease and are amenable to genetic engineering. To discern which host factors are reliable anti-coronavirus therapeutic targets, we generate mutant clonal IOs for 19 host genes previously implicated in coronavirus biology. We verify ACE2 and DPP4 as entry receptors for SARS-CoV/SARS-CoV-2 and MERS-CoV respectively. SARS-CoV-2 replication in IOs does not require the endosomal Cathepsin B/L proteases, but specifically depends on the cell surface protease TMPRSS2. Other TMPRSS family members were not essential. The newly emerging coronavirus variant B.1.1.7, as well as SARS-CoV and MERS-CoV similarly depended on TMPRSS2. These findings underscore the relevance of non-transformed human models for coronavirus research, identify TMPRSS2 as an attractive pan-coronavirus therapeutic target, and demonstrate that an organoid knockout biobank is a valuable tool to investigate the biology of current and future emerging coronaviruses., Rapid identification of host genes essential for virus replication may expedite the generation of therapeutic interventions. Here the authors generate mutant clonal intestinal organoids for 19 host genes previously implicated in coronavirus biology and identify the cell surface protease TMPRSS2 as a potential therapeutic target.

Published
2021

Catalog

Books, media, physical & digital resources
See catalog results