ArticlePloS one2019
Development of CRISPR/Cas9-mediated gene disruption systems in Giardia lamblia.
Article in PloS one, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 24 citations in OpenAlex.
- Identification of an uncharacterized protein as a novel regulator ofJournal of virology · 2025Article
- Mlf mediates proteotoxic response via formation of cellular foci for protein folding and degradation in Giardia.PLoS pathogens · 2024Article
- Harnessing the power of new genetic tools to illuminate Giardia biology and pathogenesis.Genetics · 2024Review
- A myeloid leukemia factor homolog is involved in tolerance to stresses and stress-induced protein metabolism in Giardia lamblia.Biology direct · 2023Article
- Identification of target genes regulated by encystation-induced transcription factor Myb2 using knockout mutagenesis in Giardia lamblia.Parasites & vectors · 2022Article
- Efficient CRISPR/Cas9-mediated gene disruption in the tetraploid protistOpen biology · 2022Article
- A Novel Spo11 Homologue Functions as a Positive Regulator in Cyst Differentiation inInternational journal of molecular sciences · 2021Article
- A Novel Multiprotein Bridging Factor 1-Like Protein Induces Cyst Wall Protein Gene Expression and Cyst Differentiation inInternational journal of molecular sciences · 2021Article
- CRISPR/Cas9 Ribonucleoprotein-Based Genome Editing Methodology in the Marine Protozoan ParasiteFrontiers in bioengineering and biotechnology · 2021Article
- DNA topoisomerase IIIβ promotes cyst generation by inducing cyst wall protein gene expression inOpen biology · 2020Article
- Phosphoinositide-binding proteins mark, shape and functionally modulate highly-diverged endocytic compartments in the parasitic protist Giardia lamblia.PLoS pathogens · 2020Article
- Transfection With Plasmid Causing Stable Expression of a Foreign Gene Affects General Proteome Pattern inFrontiers in cellular and infection microbiology · 2020Article
- High Cysteine Membrane Proteins (HCMPs) Are Up-Regulated DuringFrontiers in genetics · 2020Article
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Giardia lamblia becomes dormant by differentiation into a water-resistant cyst that can infect a new host. Synthesis of three cyst wall proteins (CWPs) is the fundamental feature of this differentiation. Myeloid leukemia factor (MLF) proteins are involved in cell differentiation, and tumorigenesis in mammals, but little is known about its role in protozoan parasites. We developed a CRISPR/Cas9 system to understand the role of MLF in Giardia. Due to the tetraploid genome in two nuclei of Giardia, it could be hard to disrupt a gene completely in Giardia. We only generated knockdown but not knockout mutants. We found that knockdown of the mlf gene resulted in a significant decrease of cwp gene expression and cyst formation, suggesting a positive role of MLF in encystation. We further used mlf as a model gene to improve the system. The addition of an inhibitor for NHEJ, Scr7, or combining all cassettes for gRNA and Cas9 expression into one plasmid resulted in improved gene disruption efficiencies and a significant decrease in cwp gene expression. Our results provide insights into a positive role of MLF in inducing Giardia differentiation and a useful tool for studies in Giardia.
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