Gene disruption was performed with the TALEN system. TALEN plasmids were constructed using the Platinum Gate TALEN Kit (Addgene, Cambridge, MA, USA; Kit #1000000043) as described previously[17 (link)] with modifications described as follows: the plasmid backbone of ptCMV-136/63-VR vectors were replaced with pCS2P, containing SP6 promoter for in vitro transcription. Medaka ptena and ptenb TALEN constructs were digested with NotI (Thermo Fisher Scientific, Waltham, MA, USA) and were transcribed in vitro with the use of a mMessage mMachine SP6 transcription kit (Thermo Fisher Scientific).
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