期刊
PLOS ONE
卷 14, 期 3, 页码 -出版社
PUBLIC LIBRARY SCIENCE
DOI: 10.1371/journal.pone.0213594
关键词
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资金
- Ministry of Science and Technology in Taiwan [MOST 99-2320-B-002-017-MY3, 100-2325-B-002-039, 101-2325-B-002-036-, 103-2628-B-002-006-MY3, 106-2320-B-002 -038 -MY2]
- National Health Research Institutes in Taiwan [NHRI-EX99-9510NC]
- Department of Medical Research in National Taiwan University Hospital
- Aim for the Top University Program of National Taiwan University [33474]
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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