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Genetics. 2014 May;197(1):77-89. doi: 10.1534/genetics.113.161091. Epub 2014 Mar 20.

TALENs mediate efficient and heritable mutation of endogenous genes in the marine annelid Platynereis dumerilii.

Author information

1
Max F. Perutz Laboratories, University of Vienna, 1030 Vienna, Austria.

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.

KEYWORDS:

chronobiology; evolution; genome editing; invertebrate; marine

PMID:
24653002
PMCID:
PMC4012502
DOI:
10.1534/genetics.113.161091
[Indexed for MEDLINE]
Free PMC Article

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