Implementation of a tunable t-CRISPRi system for gene regulation in Giardia duodenalis

García-Huerta, Eduardo; Espinoza-Corona, Sara; Lagunas‐Rangel, Francisco Alejandro; Ortega-Pierres, M. Guadalupe; Bermúdez‐Cruz, Rosa María

doi:10.1016/j.plasmid.2022.102641

Cited by 2 publications

(2 citation statements)

References 40 publications

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“…Through their experiments, they were able to enhance tumorigenicity in murine models by deleting SNORD50A/B using CRISPR/Cas9, which had an inhibitory effect on the Ras oncoprotein (responsible for modulating signaling pathways), promoting the uncontrolled growth of tumor cells [ 60 , 61 , 62 ]. Finally, the first studies on the modulation of gene expression in Giardia using a derivative of this technique, CRISPRi, have been carried out, demonstrating the potential of this approach as a therapeutic resource [ 63 , 64 ]. In these experiments, two distinct transcriptional repression systems were designed independently to selectively inhibit the expression of specific genes, thus compromising parasite viability [ 63 , 64 ].…”

Section: Nucleolus Of G Lambliamentioning

confidence: 99%

“…Finally, the first studies on the modulation of gene expression in Giardia using a derivative of this technique, CRISPRi, have been carried out, demonstrating the potential of this approach as a therapeutic resource [ 63 , 64 ]. In these experiments, two distinct transcriptional repression systems were designed independently to selectively inhibit the expression of specific genes, thus compromising parasite viability [ 63 , 64 ].…”

Section: Nucleolus Of G Lambliamentioning

confidence: 99%

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Analysis of Giardia lamblia Nucleolus as Drug Target: A Review

et al. 2023

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Giardia lamblia (G. lamblia) is the main causative agent of diarrhea worldwide, affecting children and adults alike; in the former, it can be lethal, and in the latter a strong cause of morbidity. Despite being considered a predominant disease in low-income and developing countries, current migratory flows have caused an increase in giardiasis cases in high-income countries. Currently, there is a wide variety of chemotherapeutic treatments to combat this parasitosis, most of which have potentially serious side effects, such as genotoxic, carcinogenic, and teratogenic. The necessity to create novel treatments and discover new therapeutic targets to fight against this illness is evident. The current review centers around the controversial nucleolus of G. lamblia, providing a historical perspective that traces its apparent absence to the present evidence supporting its existence as a subnuclear compartment in this organism. Additionally, possible examples of ncRNAs and proteins ubiquitous to the nucleolus that can be used as targets of different therapeutic strategies are discussed. Finally, some examples of drugs under research that could be effective against G. lamblia are described.

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Section: Nucleolus Of G Lambliamentioning

confidence: 99%

Section: Nucleolus Of G Lambliamentioning

confidence: 99%

Analysis of Giardia lamblia Nucleolus as Drug Target: A Review

et al. 2023

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The rise and future of CRISPR-based approaches for high-throughput genomics

Vercauteren,

Fiesack,

Maroc

et al. 2024

FEMS Microbiology Reviews

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Clustered regularly interspaced short palindromic repeats (CRISPR) has revolutionized the field of genome editing. To circumvent the permanent modifications made by traditional CRISPR techniques and facilitate the study of both essential and non-essential genes, CRISPR interference (CRISPRi) was developed. This gene silencing technique employs a deactivated Cas effector protein and a guide RNA to block transcription initiation or elongation. Continuous improvements and a better understanding of the mechanism of CRISPRi have expanded its scope, facilitating genome-wide high-throughput screens to investigate the genetic basis of phenotypes. Additionally, emerging CRISPR-based alternatives have further expanded the possibilities for genetic screening. This review delves into the mechanism of CRISPRi, compares it with other high-throughput gene-perturbation techniques and highlights its superior capacities for studying complex microbial traits. We also explore the evolution of CRISPRi, emphasizing enhancements that have increased its capabilities, including multiplexing, inducibility, titratability, predictable knockdown efficacy, and adaptability to non-model microorganisms. Beyond CRISPRi, we discuss CRISPR activation, RNA-targeting CRISPR systems and single-nucleotide resolution perturbation techniques for their potential in genome-wide high-throughput screens in microorganisms. Collectively, this review gives a comprehensive overview of the general workflow of a genome-wide CRISPRi screen, with an extensive discussion of strengths and weaknesses, future directions and potential alternatives.

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Implementation of a tunable t-CRISPRi system for gene regulation in Giardia duodenalis

Cited by 2 publications

References 40 publications

Analysis of Giardia lamblia Nucleolus as Drug Target: A Review

Analysis of Giardia lamblia Nucleolus as Drug Target: A Review

The rise and future of CRISPR-based approaches for high-throughput genomics

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