Wirdnam, Corina D; Warmus, Dawid; Faso, Carmen (2024). Nourseothricin as a novel drug for selection of transgenic Giardia lamblia. (In Press). International journal for parasitology. Drugs and drug resistance, 25(100543), p. 100543. Elsevier 10.1016/j.ijpddr.2024.100543
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Functional gene and protein characterizations in parasitic protists are often limited by their genetic tractability. Despite the development of CRISPR-Cas9-derived or inspired approaches for a handful of protist parasites, the overall genetic tractability of these organisms remains limited. The intestinal parasite Giardia lamblia is one such species, with the added challenge of a paucity of reliable selection markers. To address this limitation, we tested the feasibility of using Nourseothricin as an effective selection agent in Giardia. Here, we report that axenically-grown WB Giardia cells are sensitive to Nourseothricin and that engineering expression of the streptothricin acetyltransferase (SAT-1) gene from Streptomyces rochei in transgenic parasites confers resistance to this antibiotic. Furthermore, we determine that SAT-1-expressing parasites are cross-resistant neither to Neomycin nor Puromycin, which are widely used to select for transgenic parasites. Consequently, we show that Nourseothricin can be used in sequential combination with both Neomycin and Puromycin to select for dual transfection events. This work increases the number of reliable selection agents and markers for Giardia genetic manipulation, expanding the limited molecular toolbox for this species of global medical importance.
Item Type: |
Journal Article (Original Article) |
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Division/Institute: |
08 Faculty of Science > Department of Biology > Institute of Cell Biology 04 Faculty of Medicine > Service Sector > Institute for Infectious Diseases 05 Veterinary Medicine > Other Institutions > Centers Vetsuisse Faculty > Multidisciplinary Center for Infectious Diseases (MCID) |
Graduate School: |
Graduate School for Cellular and Biomedical Sciences (GCB) |
UniBE Contributor: |
Wirdnam, Corina-Daniela, Warmus, Dawid Adam, Faso, Carmen |
Subjects: |
500 Science > 570 Life sciences; biology 600 Technology > 610 Medicine & health |
ISSN: |
2211-3207 |
Publisher: |
Elsevier |
Language: |
English |
Submitter: |
Pubmed Import |
Date Deposited: |
30 Apr 2024 16:05 |
Last Modified: |
01 May 2024 15:24 |
Publisher DOI: |
10.1016/j.ijpddr.2024.100543 |
PubMed ID: |
38685159 |
Uncontrolled Keywords: |
Antibiotic Drug Giardia lamblia Resistance Selection marker Transfection Transgenesis |
BORIS DOI: |
10.48350/196400 |
URI: |
https://boris.unibe.ch/id/eprint/196400 |
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