WCRB2014 POSTER PRESENTATIONS (1) (335 abstracts)
Genome editing technology using ZFN and TALEN is effective on cultured porcine ST cells knockout pig
Tsubasa Kato 1 , M Sakaue 2 , S Hisamatsu 3 , M Kamoshita 1 , H Obata 1 , Tetsushi Sakuma 4 & Takashi Yamamoto 4
1Graduate School of Veterinary Sciences, Azabu University, Kanagawa, Japan; 2School of Veterinary Medicine, Azabu University, Kanagawa, Japan; 3Faculty of Life and Environmental Sciences, Azabu University, Kanagawa, Japan; 4Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Hiroshima, Japan.
In recent years, advances in molecular biology and reproductive engineering make generation of knockout (KO) animals possible. KO animals are useful and powerful tools for agriculture as well as biomedical research. Zinc finger nuclease (ZFN) and transcription activator-like effector nuclease (TALEN) genome editing technology enables site directed engineering of the genome. The aim of the present study was to ZFN and TALEN system could work on editing genome of cultured porcine Sertoli (ST) cells. DNAs of the ZFN or the TALEN were designed for introducing mutations to the genomes of GH receptor (GHR) and myostatin (MSTN). The ST cells were cultured in vitro for 24 h at 38.5 °C, 5% CO2 and saturated humidity. The transfections of DNAs of the ZFN or the TALEN into ST cells were performed by using Lipofectamine™ LTX. We examined whether the transfections were successful or not by immunostaining with anti-flag ABM2, and that gene mutations were identified by using detection kit (Cel I assay). The results of immunostaining regardless of the target in both ZFN and TALEN, showed introduction into about 10% of the ST cells was observed, and that we confirmed the mutation by using Cel I assay. Genome editing technologies using the ZFN and TALEN are effective on cultured porcine ST cells.
Article tools
My recent searches
My recently viewed abstracts
Authors
Reproduction Abstracts
ISSN 2052-1472 (online)
© Bioscientifica 2024 |
Privacy policy |
Cookie settings
BiosciAbstracts
Bioscientifica Abstracts is the gateway to a series of products that provide a permanent, citable record of abstracts for biomedical and life science conferences.
Find out more