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(P Pleurodeles Dev (ck waltl) v. Growth Di CRE-l KO) cko ffer., 2013 loxp DNA TALEN D cko TA loxp Dev. Growth ck O CR TALEN Differ., 2014 cko KO (cko) CDK1 cko RE 4 O 10 ) cko 1 cko O

loxp TALEN CRISPR CRE 1cKO CDK1 cko CDK1 TALEN loxp DNA DNA loxp 120 bp DNA TAL-Pitch vector CRE CDK1 TALEN 2CRE cko CRE IR-LEGO HSPGFP IR-LEGO GFP 3 25Gbp Inverse PCR Genomic walking RNA sequence RNA de novo sequence (4) CRISPR/CAS9 system TALEN CRISPR/Cas9 CRISPR/Cas9 system grna Cas9 mrna 1cKO TALEN PCR TALEN mrna

C CDK1 CDK K1 D DNA l loxp C CDK1 DNA TAL-Pitch vector loxp DNA DN NA TA AL-Pitch vector loxp cko 2 IR-LEGO GF FP IR-LEGO Temperatu ure stimulater cko HS SP-CRE HSP P CRE-lo oxp H HSP CRE 3 3 CDK1 cyclin D1 cyclin D1 cyclin D1 4 4CR RISPR/Cas9 system RN NA guiderna Cas s9 mrna g guide RNA TALEN

KO 5 (2016) 67, 264-269. - (2016) 34, 1473-1476. Tane, S., Okayama, H., Ikenishi, A., Amemiya, Y., Nakayama, I. K., and Takeuchi, T*. Two inhibitory systems and CKIs regulate cell cycle exit of mammalian cardiomyocytes after birth. Biochem. Biophys. Res. Commun., (2015) 466, 147-154. doi: 10.1016/j.bbrc.2015.08.102. Kawasumi, A., Hayashi, T., Kobayashi, T., Nagayama, C., Hayashi, S., Kamei, Y., Morishita, Y., T Takeuchi, T., Tamura, K., Yokoyama, H*. Application of local gene induction by infrared laser-mediated microscope and temperature stimulator to amphibian regeneration study. Dev. Growth Differ.,(2015) 57, 601-613. doi: 10.1111/dgd.12241. Tane, S., Kubota, M., Okayama, H., Ikenishi, A., Yoshitome, S., Iwamoto, N., Satoh, Y., Kusakabe, A., Ogawa, S., Kanai, A., Molkentin, J. D., Nakamura, K., Ohbayashi, T., and Takeuchi, T*. Repression of cyclin D1 expression is necessary for the maintenance of cell cycle exit in adult mammalian cardiomyocytes. J. Biol. Chem., (2014) 289, 18033-18044. doi: 10.1074/jbc.M113.541953. 17 2017 3 8 Hayashi T., Myouga, A., Tsuchiya, E., Azuma, S., Sato and Takeuchi, T.Cardiac regeneration in newts achieved based on the compensatory manner. ICZ/ZSJ Joint Meeting Symposium (invited) Nov 17 th, 2017 OIST, Naha city, Okinawa CREST 2017 2 20 2015 12 1 2015 12 1 2015 12 3,4 2015 9 18 Takeuchi, T., Tane, S., Okayama, Ikenishi, A., H., Amemiya Y., Ohdira, Y., Myouga, A., Azuma, S., Hayashi, T. What determines differences in cardiac regenerative abilities between mouse and newts? 2014 11 25-27 cyclin D1 2014 11 25-27 2014 11 25-27

2014 11 25-27 2014 11 25-27 Hayashi, T., Sakuma, T., Myouga, A., Sakamoto, K., Yokotani, N., Inoue, T., Kawaguchi, E., Agata, K., Yamamoto, T., and Takeuchi, T. Genome editing is a powerful tool for studies in newt cardiac regeneration. 2014917 Takeuchi, T., Tane, S., Okayama, Ikenishi, A., Amemiya, Y., Myouga, A., Hayashi, T. Mechanisms of cell cycle exit and the maintenance in mouse cardiomyocytes, and the roles for regenerative abilities in mouse and newts. EMBO Conference The molecular &Cellular Basis of Regeneration & Tissue Repair, Sept. 6-10, 2014, Sant Feliu de Guixols (Spain). Hayashi, T., Myouga, A., Yokotani, N., Takeuchi, T. Development of molecular genetic system for newts and linage tracing analyses in cardiac regenerarion. EMBO Conference The molecular &Cellular Basis of Regeneration & Tissue Repair, Sept. 6-10, 2014, Sant Feliu de Guixols (Spain). Tane, S., Okayama, H., Ikenishi, A., Satoh, Y., Takeuchi, T. Cell cycle exit in cardiomyocytes is initiated by p21 Cip1 and p27 Kip1, and maintained by two inhibitor systems. 2014 5 27-30 Hayashi, T., Sakamoto, K., Sakuma, T., Myouga, A., Yokotani, N., Inoue, T., Kawaguchi, E, Agata, K., Yamamoto, T., Takeuchi, T. TALEN-mediated genome editing is very useful in Iberian ribbed newts (Pleurodeles waltl), an experimental model animal for regeneration. 2014 5 27-30 4 (2017) (2017) Hayashi, T., and Takeuchi, T. (2015) Mutagenesis in Newts: Protocol for Iberian Ribbed Newts. In TALENs: Methods and Protocols. (eds. Kuhn, R., Wefers, B., and Wurst, W.), pp119-126, Springer New York. doi: 10.1007/978-1-4939-2932-0_10. Hayashi, T. and Takeuchi, T. (2015) Gene Manipulation for Regenerative Studies Using the Iberian Ribbed Newt, Pleurodeles waltl. In Salamanders in Regeneration Research. (eds. A. Kumar and A. Simon), pp297-305, Springer New York. doi: 10.1007/978-1-4939-2495-0_23. 60580925