title: Use of Translation Blocking Morpholinos for Gene Knockdown in Giardia lamblia.
abstract: Giardia lamblia, a major parasite, is an emerging model organism due to its compact genomic arrangement and composition. The most popular reverse genetic technique, RNAi, is ineffective in Giardia. In contrast, protein depletion by translation blocking morpholinos is suitable for most gene targets and provides up to 80% depletion of the target protein. The method is fast, reliable, and specific. After antisense morpholino oligomer delivery into Giardia trophozoites by electroporation, the cells can be used for many subsequent analyses 8-48 h after treatment. In this chapter, suitable gene tags, plasmids, and techniques necessary for proper morpholino targeting are described.
authors.affiliation: Department of Biology, University of Washington, Seattle, WA, 98195, USA.|Department of Experimental Plant Biology, Faculty of Science, Charles University in Prague, Viničná 5, 128 44, Prague 2, Czech Republic.
authors.forename: Jana
authors.initials: J
authors.lastname: Krtková
authors.affiliation: Department of Biology, University of Washington, Seattle, WA, 98195, USA. aparedez@uw.edu.
authors.forename: Alexander R
authors.initials: AR
authors.lastname: Paredez
mesh_terms: D001483:Base Sequence; D003001:Cloning, Molecular; D018274:Electroporation; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D023061:Gene Order; D005822:Genetic Vectors / Q000235:genetics; D016829:Giardia lamblia / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D060172:Morpholinos / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics*; D014176:Protein Biosynthesis / Q000235:genetics*; D015800:Protozoan Proteins / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins; D014170:Transformation, Genetic
chemical_list: D060172:Morpholinos; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins
keywords: Giardia; Integration; Knockdown; Morpholino; Quantitative Western blotting