title: Validation of Protein Knockout in Mutant Zebrafish Lines Using In Vitro Translation Assays.
abstract: Advances in genome-editing technology have made creation of zebrafish mutant lines accessible to the community. Experimental validation of protein knockout is a critical step in verifying null mutants, but this can be a difficult task. Absence of protein can be confirmed by Western blotting; however, this approach requires target-specific antibodies that are generally not available for zebrafish proteins. We address this issue using in vitro translation assays, a fast and standard procedure that can be easily implemented.
authors.affiliation: 1 Whitehead Institute for Biomedical Research , Cambridge, Massachusetts.
authors.forename: Bradley S
authors.initials: BS
authors.lastname: Carter
authors.affiliation: 1 Whitehead Institute for Biomedical Research , Cambridge, Massachusetts.
authors.forename: Christian
authors.initials: C
authors.lastname: Cortés-Campos
authors.affiliation: 2 Department of Biology, Massachusetts Institute of Technology , Cambridge, Massachusetts.
authors.forename: Xiao
authors.initials: X
authors.lastname: Chen
authors.affiliation: 1 Whitehead Institute for Biomedical Research , Cambridge, Massachusetts.
authors.forename: Jasmine M
authors.initials: JM
authors.lastname: McCammon
authors.affiliation: 1 Whitehead Institute for Biomedical Research , Cambridge, Massachusetts.|2 Department of Biology, Massachusetts Institute of Technology , Cambridge, Massachusetts.
authors.forename: Hazel L
authors.initials: HL
authors.lastname: Sive
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics / Q000378:metabolism*; D002474:Cell-Free System; D000072669:Gene Editing; D055786:Gene Knockout Techniques / Q000379:methods*; D009154:Mutation; D014176:Protein Biosynthesis; D015027:Zebrafish / Q000235:genetics / Q000378:metabolism*; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D029961:Zebrafish Proteins
keywords: in vitro translation; mutant; zebrafish