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Transcription activator‐like effector nucleases efficiently disrupt the target gene in Iberian ribbed newts ( P leurodeles waltl ), an experimental model animal for regeneration
Author(s) -
Hayashi Toshinori,
Sakamoto Kousuke,
Sakuma Tetsushi,
Yokotani Naoki,
Inoue Takeshi,
Kawaguchi Eri,
Agata Kiyokazu,
Yamamoto Takashi,
Takeuchi Takashi
Publication year - 2014
Publication title -
development, growth and differentiation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.864
H-Index - 66
eISSN - 1440-169X
pISSN - 0012-1592
DOI - 10.1111/dgd.12103
Subject(s) - transcription activator like effector nuclease , biology , effector , genome editing , gene knockout , gene , regeneration (biology) , genetics , gene targeting , model organism , genome , microbiology and biotechnology , computational biology
Regeneration of a lost tissue in an animal is an important issue. Although regenerative studies have a history of research spanning more than a century, the gene functions underlying regulation of the regeneration are mostly unclear. Analysis of knockout animals is a very powerful tool with which to elucidate gene function. Recently, transcription activator‐like effector nucleases ( TALEN s) have been developed as an effective technique for genome editing. This technique enables gene targeting in amphibians such as newts that were previously impossible. Here we show that newts microinjected with TALEN m RNA s designed for targeting the tyrosinase gene in single‐cell stage embryos revealed an albino phenotype. Sequence analysis revealed that the tyrosinase genes were effectively disrupted in these albino newts. Moreover, precise genome alteration was achieved using TALEN s and single strand oligodeoxyribonucleotides. Our results suggest that TALEN s are powerful tools for genome editing for regenerative research in newts.