ArticleMolecular biology of the cell2019
Robust and stable transcriptional repression in Giardia using CRISPRi.
Article in Molecular biology of the cell, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 33 papers.
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Who cites it
33 citing papers in PubMed, 69 citations in OpenAlex.
- Arginine metabolism has a pivotal function for the encystation of Giardia duodenalis.PLoS pathogens · 2026Article
- Identification of an uncharacterized protein as a novel regulator ofJournal of virology · 2025Article
- Two Centrins and Their Posttranslational Modification Modulate the Cell Cycle of Giardia lamblia.MicrobiologyOpen · 2025Article
- Article
- Giardia intraflagellar transport protein 88 is involved in flagella formation.Parasites, hosts and diseases · 2025Article
- Unraveling the role of cAMP signaling inmSphere · 2024Article
- Malate dehydrogenase in parasitic protozoans: roles in metabolism and potential therapeutic applications.Essays in biochemistry · 2024Review
- The rise and future of CRISPR-based approaches for high-throughput genomics.FEMS microbiology reviews · 2024Review
- Microbial Matryoshka: Addressing the Relationship between Pathogenic Flagellated Protozoans and Their RNA Viral Endosymbionts (FamilyVeterinary sciences · 2024Review
- Harnessing the power of new genetic tools to illuminate Giardia biology and pathogenesis.Genetics · 2024Review
- Encystation stimuli sensing is mediated by adenylate cyclase AC2-dependent cAMP signaling in Giardia.Nature communications · 2023Article
- Spliceosomal introns in the diplomonad parasiteMicrobial genomics · 2023Article
- Analysis ofPharmaceuticals (Basel, Switzerland) · 2023Review
- Identification of target genes regulated by encystation-induced transcription factor Myb2 using knockout mutagenesis in Giardia lamblia.Parasites & vectors · 2022Article
- Kinesin-13, a Motor Protein, is Regulated by Polo-like Kinase in Giardia lamblia.The Korean journal of parasitology · 2022Article
- A cell-cycle-dependent GARP-like transcriptional repressor regulates the initiation of differentiation inProceedings of the National Academy of Sciences of the United States of America · 2022Article
- Efficient CRISPR/Cas9-mediated gene disruption in the tetraploid protistOpen biology · 2022Article
- Disc and Actin Associated Protein 1 influences attachment in the intestinal parasite Giardia lamblia.PLoS pathogens · 2022Article
- CRISPR Interference as a Tool to Repress Gene Expression in Haloferax volcanii.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Functional Identification of a Nuclear Localization Signal of MYB2 Protein in Giardia lamblia.The Korean journal of parasitology · 2020Article
Corrections and comments
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Authors and funding
8 authors at 1 institution in 1 country.
Funding
Abstract
Giardia lamblia is a binucleate protistan parasite causing significant diarrheal disease worldwide. An inability to target Cas9 to both nuclei, combined with the lack of nonhomologous end joining and markers for positive selection, has stalled the adaptation of CRISPR/Cas9-mediated genetic tools for this widespread parasite. CRISPR interference (CRISPRi) is a modification of the CRISPR/Cas9 system that directs catalytically inactive Cas9 (dCas9) to target loci for stable transcriptional repression. Using a Giardia nuclear localization signal to target dCas9 to both nuclei, we developed efficient and stable CRISPRi-mediated transcriptional repression of exogenous and endogenous genes in Giardia. Specifically, CRISPRi knockdown of kinesin-2a and kinesin-13 causes severe flagellar length defects that mirror defects with morpholino knockdown. Knockdown of the ventral disk MBP protein also causes severe structural defects that are highly prevalent and persist in the population more than 5 d longer than defects associated with transient morpholino-based knockdown. By expressing two guide RNAs in tandem to simultaneously knock down kinesin-13 and MBP, we created a stable dual knockdown strain with both flagellar length and disk defects. The efficiency and simplicity of CRISPRi in polyploid Giardia allows rapid evaluation of knockdown phenotypes and highlights the utility of CRISPRi for emerging model systems.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.