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Double diploid (essentially tetraploid) Giardia trophozoites have presented a formidable challenge to the development of molecular genetic tools to interrogate gene function. High sequence divergence and the high percentage of hypothetical proteins lacking homology to proteins in other eukaryotes have limited our understanding of Giardia protein function, slowing drug target validation and development. For more than 25 years, Giardia A and B assemblages have been readily amenable to transfection with plasmids or linear DNA templates. Here, we highlight the utility and power of genetic approaches developed to assess protein function in Giardia, with particular emphasis on the more recent clustered regularly interspaced palindromic repeats\/Cas9-based methods for knockdowns and knockouts. Robust and reliable molecular genetic approaches are fundamental toward the interrogation of Giardia protein function and evaluation of druggable targets. New genetic approaches tailored for the double diploid Giardia are imperative for understanding Giardia's unique biology and pathogenesis.<\/jats:p>","DOI":"10.1093\/genetics\/iyae038","type":"journal-article","created":{"date-parts":[[2024,4,16]],"date-time":"2024-04-16T19:57:18Z","timestamp":1713297438000},"source":"Crossref","is-referenced-by-count":8,"title":["Harnessing the power of new genetic tools to illuminate <i>Giardia<\/i> biology and pathogenesis"],"prefix":"10.1093","volume":"227","author":[{"given":"Kari D","family":"Hagen","sequence":"first","affiliation":[{"name":"Department of Microbiology and Molecular Genetics, University of California , Davis, Davis, CA 95616 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christopher J S","family":"Hart","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Molecular Genetics, University of California , Davis, Davis, CA 95616 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shane G","family":"McInally","sequence":"additional","affiliation":[{"name":"Department of Biology and Biotechnology, Worcester Polytechnic Institute , Worcester, MA 01609 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0843-1759","authenticated-orcid":false,"given":"Scott C","family":"Dawson","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Molecular Genetics, University of California , Davis, Davis, CA 95616 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2024,4,16]]},"reference":[{"issue":"12","key":"2024061207404779900_iyae038-B1","doi-asserted-by":"publisher","first-page":"2498","DOI":"10.1093\/gbe\/evt197","article-title":"Genome sequencing of Giardia lamblia genotypes A2 and B isolates (DH and GS) and comparative analysis with the genomes of genotypes A1 and E (WB 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