title: TALENs mediate efficient and heritable mutation of endogenous genes in the marine annelid Platynereis dumerilii.
abstract: Platynereis dumerilii is a marine polychaete and an established model system for studies of evolution and development. Platynereis is also a re-emerging model for studying the molecular basis of circalunar reproductive timing: a biological phenomenon observed in many marine species. While gene expression studies have provided new insight into patterns of gene regulation, a lack of reverse genetic tools has so far limited the depth of functional analyses in this species. To address this need, we established customized transcriptional activator-like effector nucleases (TALENs) as a tool to engineer targeted modifications in Platynereis genes. By adapting a workflow of TALEN construction protocols and mutation screening approaches for use in Platynereis, we engineered frameshift mutations in three endogenous Platynereis genes. We confirmed that such mutations are heritable, demonstrating that TALENs can be used to generate homozygous knockout lines in P. dumerilii. This is the first use of TALENs for generating genetic knockout mutations in an annelid model. These tools not only open the door for detailed in vivo functional analyses, but also can facilitate further technical development, such as targeted genome editing.
authors.affiliation: Max F. Perutz Laboratories, University of Vienna, 1030 Vienna, Austria.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Bannister
authors.forename: Olga
authors.initials: O
authors.lastname: Antonova
authors.forename: Alessandra
authors.initials: A
authors.lastname: Polo
authors.forename: Claudia
authors.initials: C
authors.lastname: Lohs
authors.forename: Natalia
authors.initials: N
authors.lastname: Hallay
authors.forename: Agne
authors.initials: A
authors.lastname: Valinciute
authors.forename: Florian
authors.initials: F
authors.lastname: Raible
authors.forename: Kristin
authors.initials: K
authors.lastname: Tessmar-Raible
mesh_terms: D000818:Animals; D001483:Base Sequence; D053837:DNA Cleavage; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D005818:Genetic Engineering / Q000379:methods*; D060005:Genotyping Techniques; D007814:Larva / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D011077:Polychaeta / Q000235:genetics*; D015203:Reproducibility of Results; D017384:Sequence Deletion; D015053:Zygote / Q000378:metabolism
chemical_list: D004262:DNA Restriction Enzymes
keywords: chronobiology; evolution; genome editing; invertebrate; marine