University of Tsukuba 3 articles published in JoVE Genetics Primordial Germ Cell Cryopreservation and Revival of Drosophila Strains Kaori Nishimura*1, Miho Asaoka*2, Yurina Sakamaki*2, Tatsuya Fukumoto*3,4, Daisuke Tanaka3, Satoru Kobayashi2, Toshiyuki Takano-Shimizu-Kouno1 1KYOTO Drosophila Stock Center, Kyoto Institute of Technology, 2Life Science Center for Survival Dynamics, Tsukuba Advanced Research Alliance (TARA), University of Tsukuba, 3Research Center of Genetic Resources, National Agriculture and Food Research Organization, 4Shizuoka Prefectural Ogasa High School A long-term preservation method for Drosophila strains as an alternative to the frequent transfer of adult flies to fresh food vials is highly desirable. This protocol describes the cryopreservation of Drosophila primordial germ cells and strain revival via their transplantation to agametic host embryos. Developmental Biology Zygote Microinjection for Creating Gene Cassette Knock-in and Flox Alleles in Mice Yoko Tanimoto*1, Natsuki Mikami*1,2, Miyuki Ishida1, Natsumi Iki1, Kanako Kato1, Fumihiro Sugiyama1, Satoru Takahashi1, Seiya Mizuno1 1Laboratory Animal Resource Center and Trans-Border Medical Research Center, University of Tsukuba, 2Ph.D. Program in Human Biology, School of Integrative and Global Majors, University of Tsukuba The present protocol describes zygote microinjection of CRISPR-Cas9 and donor DNA to efficiently produce gene cassette knock-in and floxed mice. Biology Isolation and Culture of Primary Oral Keratinocytes from the Adult Mouse Palate Yen Xuan Ngo1,2,3, Kenta Haga4, Ayako Suzuki4, Hiroko Kato4, Hiromi Yanagisawa1,5, Kenji Izumi4, Aiko Sada1,3 1Life Science Center for Survival Dynamics, Tsukuba Advanced Research Alliance (TARA), University of Tsukuba, 2Ph.D. Program in Human Biology, School of Integrative and Global Majors, University of Tsukuba, 3International Research Center for Medical Sciences (IRCMS), Kumamoto University, 4Division of Biomimetics, Faculty of Dentistry and Graduate School of Medical and Dental Sciences, Niigata University, 5Faculty of Medicine, University of Tsukuba The present protocol describes the isolation and culture of oral keratinocytes derived from the adult mouse palate. An evaluation method using immunostaining is also reported.
