A5053259162- Trypanosoma species research and implications
- Research on Leishmaniasis Studies
- CRISPR and Genetic Engineering
- SARS-CoV-2 and COVID-19 Research
- COVID-19 Clinical Research Studies
- Microtubule and mitosis dynamics
- Insect symbiosis and bacterial influences
- Genetic and Kidney Cyst Diseases
- Micro and Nano Robotics
- Plant Virus Research Studies
- SARS-CoV-2 detection and testing
- Photosynthetic Processes and Mechanisms
- Lysosomal Storage Disorders Research
- Innovation and Socioeconomic Development
- Parasitic Diseases Research and Treatment
- Renal Diseases and Glomerulopathies
- Polyomavirus and related diseases
- RNA Interference and Gene Delivery
- Toxin Mechanisms and Immunotoxins
- Viral gastroenteritis research and epidemiology
- Aeolian processes and effects
- Coastal wetland ecosystem dynamics
- Viral Infections and Immunology Research
- Respiratory viral infections research
- Parasites and Host Interactions
University of Oxford
2017-2025
University of Würzburg
2022-2025
Washington University in St. Louis
2022-2023
DKFZ-ZMBH Alliance
2023
University of Glasgow
2022-2023
Google (United States)
2017
Deutsche Forschungsgemeinschaft
1977
Clustered regularly interspaced short palindromic repeats (CRISPR), CRISPR-associated gene 9 (Cas9) genome editing is set to revolutionize genetic manipulation of pathogens, including kinetoplastids. CRISPR technology provides the opportunity develop scalable methods for high-throughput production mutant phenotypes. Here, we report development a CRISPR-Cas9 toolkit that allows rapid tagging and knockout in diverse kinetoplastid species without requiring user perform any DNA cloning. We...
The protozoan parasite Leishmania possesses a single flagellum, which is remodelled during the parasite's life cycle from long motile flagellum in promastigote forms sand fly to short immotile amastigotes residing mammalian phagocytes. This study examined protein composition and vivo function of flagellum. Protein mass spectrometry label free enrichment testing isolated flagella deflagellated cell bodies defined flagellar proteome for L. mexicana (available via ProteomeXchange with...
<ns4:p><ns4:bold>Background: </ns4:bold>Laboratory diagnosis of SARS-CoV-2 infection (the cause COVID-19) uses PCR to detect viral RNA (vRNA) in respiratory samples. has also been detected other sample types, but there is limited understanding the clinical or laboratory significance its detection blood.</ns4:p><ns4:p> <ns4:bold>Methods: </ns4:bold>We undertook a systematic literature review assimilate evidence for frequency vRNA blood, and identify associated characteristics. We performed...
<ns4:p><ns4:bold>Background: </ns4:bold>Laboratory diagnosis of SARS-CoV-2 infection (the cause COVID-19) uses PCR to detect viral RNA (vRNA) in respiratory samples. has also been detected other sample types, but there is limited understanding the clinical or laboratory significance its detection blood.</ns4:p><ns4:p> <ns4:bold>Methods: </ns4:bold>We undertook a systematic literature review assimilate evidence for frequency vRNA blood, and identify associated characteristics. We performed...
Background Infection with Trypanosoma cruzi causes Chagas disease, a major public health problem throughout Latin America. There is no vaccine and the only drugs have severe side effects. Efforts to generate new therapies are hampered by limitations in our understanding of parasite biology disease pathogenesis. Studies compromised complexity long-term nature infection, fact that parasites barely detectable during chronic stage. In addition, functional dissection T. has been restricted...
Abstract For the protozoan parasite Leishmania , completion of its life cycle requires sequential adaptation cellular physiology and nutrient scavenging mechanisms to different environments a sand fly alimentary tract acidic mammalian host cell phagolysosome. Transmembrane transporters are gatekeepers intracellular environments, controlling flux solutes ions across membranes. To discover which vital for survival as amastigote forms, we carried out systematic loss-of-function screen L....
CRISPR/Cas9 gene editing has revolutionised loss-of-function experiments in Leishmania , the causative agent of leishmaniasis. As lack a functional non-homologous DNA end joining pathway however, obtaining null mutants typically requires additional donor DNA, selection drug resistance-associated edits or time-consuming isolation clones. Genome-wide screens across different conditions and multiple species are therefore unfeasible at present. Here, we report cytosine base editor (CBE) toolbox...
The recent adaptation of CRISPR Cas9 genome editing to Leishmania spp. has opened a new era in deciphering biology. method was recently improved using PCR-based approach, which eliminated the need for cloning. This allows high-throughput gene deletion, successfully validated L. mexicana and major. In this study, we toolkit donovani targeting flagellar protein PF16, confirming that tagged localizes flagellum null mutants lose their motility. We then used technique characterise CK1.1, member...
The ability to analyse the function of all genes in a genome is highly desirable, yet challenging Leishmania due repetitive genome, limited DNA repair mechanisms and lack RNA interference most species. While our introduction cytosine base editor (CBE) demonstrated potential overcome these limitations (Engstler Beneke (2023)), challenges remained, including low transfection efficiency, variable editing rates across species, parasite growth effects, competition between deleterious...
Parasites of the Leishmania donovani complex are responsible for visceral leishmaniasis, a vector-borne disease transmitted through bite female phlebotomine sand flies. As well as human hosts, these parasites infect many mammals which can serve reservoirs. Dogs particularly important reservoirs in Europe. Transmission is widespread across Asia, Africa, Americas, and Mediterranean basin, including South France. Visceral leishmaniasis poses fatal threat if left untreated. Research into...
The movement and pathogenicity of trypanosomatid species, the causative agents trypanosomiasis leishmaniasis, are dependent on a flagellum that contains an axoneme dynein-bound doublet microtubules (DMTs). In this work, we present cryo–electron microscopy structures DMTs from two Leishmania tarentolae Crithidia fasciculata , at resolutions up to 2.7 angstrom. revealed 27 trypanosomatid-specific microtubule inner proteins, specialized dynein-docking complex, presence paralogous proteins...
ABSTRACT Background Laboratory diagnosis of SARS-CoV-2 infection (the cause COVID-19) uses PCR to detect viral RNA (vRNA) in respiratory samples. has also been detected other sample types, but there is limited understanding the clinical or laboratory significance its detection blood. Methods We undertook a systematic literature review assimilate evidence for frequency vRNA blood, and identify associated characteristics. performed RT-PCR serum samples from UK cohort acute convalescent...
The ability to analyze the function of all genes in a genome is highly desirable, yet challenging Leishmania due repetitive genome, limited DNA repair mechanisms, and lack RNA interference most species. While our introduction cytosine base editor (CBE) demonstrated potential overcome these limitations (Engstler Beneke, 2023), challenges remained, including low transfection efficiency, variable editing rates across species, parasite growth effects, competition between deleterious...
SUMMARY Iodination of the surface poliovirus and its artificial empty capsid demonstrated predominant labelling polypeptide VP 1 on intact particle an increased 2 capsid.
Leishmania braziliensis is the main causative agent of Tegumentary Leishmaniasis in Americas. However, difficulties related to genome manipulation, experimental infection, and parasite growth have so far limited studies with this species. CRISPR-Cas9-based technology has made editing more accessible, here we successfully employed LeishGEdit approach attenuate L. braziliensis. We generated a transgenic cell line expressing Cas9 T7 RNA polymerase, which was for targeted deletion centrin,...
Knockout phenotypes in Leishmania and detection of ULK4 ciliary localization mice show that Fused/STK36 interact a conserved pathway for stable assembly motile cilia. This study describes severe structural defects flagella resulting from the loss or shows evidence BioID co-immunoprecipitation direct interaction with pseudokinase ULK4.
Protists of the order Kinetoplastida possess a single multifunctional flagellum, which powers cellular displacement and mediates attachment to tissues arthropod vector. The kinetoplastid flagellar cytoskeleton consists nine-microtubule doublet axoneme; further structural elaborations, can vary between species life cycle stages, include assembly axonemal dynein complexes, pair singlet microtubules extra-axonemal paraflagellar rod. intracellular amastigote forms Leishmania spp. build short,...
Until 2015, loss-of-function studies to elucidate protein function in Leishmania relied on gene disruption through homologous recombination. Then, the CRISPR/Cas9 revolution reached these protozoan parasites allowing efficient genome editing with one round of transfection. In addition, development LeishGEdit, a PCR-based toolkit for generating knockouts and tagged lines using CRISPR/Cas9, allowed more straightforward effective editing. this system, plasmid pTB007 is delivered episomal...
<ns3:p>We present the genome sequence of <ns3:italic>Leishmania mexicana</ns3:italic> MNYC/BZ/62/M379 modified to express Cas9 and T7 RNA-polymerase, revealing high similarity reference (MHOM/GT2001/U1103). Through RNAseq-based annotation coding sequences untranslated regions, we provide primer for construct sgRNA template generation CRISPR-assisted gene deletion endogenous tagging.</ns3:p>
The mitochondrial signature glycerophospholipid, cardiolipin (CL), binds to transporters of the inner membrane and plays a central role in formation stability respiratory supercomplexes. Functional structural requirement CL for proteins has been studied vitro using purified reconstituted or synthesis knockout cells that are viable under specific growth conditions. However, no information is available on function, protein stability, expression levels during depletion. In contrast yeast...