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. 2019 May 14;8(5):453. doi: 10.3390/cells8050453

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© 2019 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Generation of pyrroline-5-carboxylate reductase 1 (pycr1) gene knockout (KO) zebrafish by Transcription activator-like effector nuclease (TALEN) genome editing tool. (A) Scheme showed the TALEN left and right arm sequences used to target pycr1 gene on exon 4 in zebrafish (top panel). The Left and right arm and spacer sequences used for TALEN design were listed in the middle panel. The high-resolution melting assay (HRMA) of F0 fish were showed in the bottom panel. (B) Comparison of the Sanger sequencing results of F1 generation that carried potential mutations for pycr1 gene. (C) The stable pycr1 KO fish used in this study carrying a 2 nucleotide-deletion (delCT) results in a predicted truncate PYCR1 protein with 54 amino acids. (D) The predicted three-dimensional structure of PYCR1 protein for WT (+/+) and pycr1 KO (−/−) fish.