OBJECTIVES: In order to create a suitable model for high-throughput drug screening, a Giardia lamblia WB C6 strain expressing Escherichia coli glucuronidase A (GusA) was created and tested with respect to susceptibility to the anti-giardial drugs nitazoxanide and metronidazole. METHODS: GusA, a well-established reporter gene in other systems, was cloned into the vector pPacVInteg allowing stable expression in G. lamblia under control of the promoter from the glutamate dehydrogenase (gdh) gene. The resulting transgenic strain was compared with the wild-type strain in a vitality assay, characterized with respect to susceptibility to nitazoxanide, metronidazole and -- as assessed in a 96-well plate format -- to a panel of 15 other compounds to be tested for anti-giardial activity. RESULTS: GusA was stably expressed in G. lamblia. Using a simple glucuronidase assay protocol, drug efficacy tests yielded results similar to those from cell counting. CONCLUSIONS: G. lamblia WB C6 GusA is a suitable tool for high-throughput anti-giardial drug screening.
OBJECTIVES: In order to create a suitable model for high-throughput drug screening, a Giardia lamblia WB C6 strain expressing Escherichia coli glucuronidase A (GusA) was created and tested with respect to susceptibility to the anti-giardial drugs nitazoxanide and metronidazole. METHODS: GusA, a well-established reporter gene in other systems, was cloned into the vector pPacVInteg allowing stable expression in G. lamblia under control of the promoter from the glutamate dehydrogenase (gdh) gene. The resulting transgenic strain was compared with the wild-type strain in a vitality assay, characterized with respect to susceptibility to nitazoxanide, metronidazole and -- as assessed in a 96-well plate format -- to a panel of 15 other compounds to be tested for anti-giardial activity. RESULTS: GusA was stably expressed in G. lamblia. Using a simple glucuronidase assay protocol, drug efficacy tests yielded results similar to those from cell counting. CONCLUSIONS: G. lamblia WB C6 GusA is a suitable tool for high-throughput anti-giardial drug screening.
Authors: Samantha A Michaels; Han-Wei Shih; Bailin Zhang; Edelmar D Navaluna; Zhongsheng Zhang; Ranae M Ranade; J Robert Gillespie; Ethan A Merritt; Erkang Fan; Frederick S Buckner; Alexander R Paredez; Kayode K Ojo Journal: J Antimicrob Chemother Date: 2020-05-01 Impact factor: 5.790
Authors: Christopher J S Hart; Taylah Munro; Katherine T Andrews; John H Ryan; Andrew G Riches; Tina S Skinner-Adams Journal: Int J Parasitol Drugs Drug Resist Date: 2017-01-27 Impact factor: 4.077
Authors: Samantha A Michaels; Matthew A Hulverson; Grant R Whitman; Linh T Tran; Ryan Choi; Erkang Fan; Case W McNamara; Melissa S Love; Kayode K Ojo Journal: Antimicrob Agents Chemother Date: 2022-06-15 Impact factor: 5.938
Authors: Joachim Müller; Samuel Rout; David Leitsch; Jathana Vaithilingam; Adrian Hehl; Norbert Müller Journal: Int J Parasitol Drugs Drug Resist Date: 2015-03-25 Impact factor: 4.077
Authors: Kelly M Hennessey; Ilse C Rogiers; Han-Wei Shih; Matthew A Hulverson; Ryan Choi; Molly C McCloskey; Grant R Whitman; Lynn K Barrett; Ethan A Merritt; Alexander R Paredez; Kayode K Ojo Journal: PLoS Negl Trop Dis Date: 2018-08-06