title: Site-specific recombination in the chicken genome using Flipase recombinase-mediated cassette exchange.
abstract: Targeted genome recombination has been applied in diverse research fields and has a wide range of possible applications. In particular, the discovery of specific loci in the genome that support robust and ubiquitous expression of integrated genes and the development of genome-editing technology have facilitated rapid advances in various scientific areas. In this study, we produced transgenic (TG) chickens that can induce recombinase-mediated gene cassette exchange (RMCE), one of the site-specific recombination technologies, and confirmed RMCE in TG chicken-derived cells. As a result, we established TG chicken lines that have, Flipase (Flp) recognition target (FRT) pairs in the chicken genome, mediated by piggyBac transposition. The transgene integration patterns were diverse in each TG chicken line, and the integration diversity resulted in diverse levels of expression of exogenous genes in each tissue of the TG chickens. In addition, the replaced gene cassette was expressed successfully and maintained by RMCE in the FRT predominant loci of TG chicken-derived cells. These results indicate that targeted genome recombination technology with RMCE could be adaptable to TG chicken models and that the technology would be applicable to specific gene regulation by cis-element insertion and customized expression of functional proteins at predicted levels without epigenetic influence.
authors.affiliation: *Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National University, Seoul, Korea; Graduate School of International Agricultural Technology, and Institute of Green-Bio Science and Technology, Seoul National University, Pyeongchang-gun, Gangwon-do, Korea; and Institute for Biomedical Sciences, Shinshu University, Kamiina, Nagano, Japan.
authors.forename: Hong Jo
authors.initials: HJ
authors.lastname: Lee
authors.affiliation: *Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National University, Seoul, Korea; Graduate School of International Agricultural Technology, and Institute of Green-Bio Science and Technology, Seoul National University, Pyeongchang-gun, Gangwon-do, Korea; and Institute for Biomedical Sciences, Shinshu University, Kamiina, Nagano, Japan.
authors.forename: Hyung Chul
authors.initials: HC
authors.lastname: Lee
authors.affiliation: *Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National University, Seoul, Korea; Graduate School of International Agricultural Technology, and Institute of Green-Bio Science and Technology, Seoul National University, Pyeongchang-gun, Gangwon-do, Korea; and Institute for Biomedical Sciences, Shinshu University, Kamiina, Nagano, Japan.
authors.forename: Young Min
authors.initials: YM
authors.lastname: Kim
authors.affiliation: *Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National University, Seoul, Korea; Graduate School of International Agricultural Technology, and Institute of Green-Bio Science and Technology, Seoul National University, Pyeongchang-gun, Gangwon-do, Korea; and Institute for Biomedical Sciences, Shinshu University, Kamiina, Nagano, Japan.
authors.forename: Young Sun
authors.initials: YS
authors.lastname: Hwang
authors.affiliation: *Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National University, Seoul, Korea; Graduate School of International Agricultural Technology, and Institute of Green-Bio Science and Technology, Seoul National University, Pyeongchang-gun, Gangwon-do, Korea; and Institute for Biomedical Sciences, Shinshu University, Kamiina, Nagano, Japan.
authors.forename: Young Hyun
authors.initials: YH
authors.lastname: Park
authors.affiliation: *Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National University, Seoul, Korea; Graduate School of International Agricultural Technology, and Institute of Green-Bio Science and Technology, Seoul National University, Pyeongchang-gun, Gangwon-do, Korea; and Institute for Biomedical Sciences, Shinshu University, Kamiina, Nagano, Japan.
authors.forename: Tae Sub
authors.initials: TS
authors.lastname: Park
authors.affiliation: *Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National University, Seoul, Korea; Graduate School of International Agricultural Technology, and Institute of Green-Bio Science and Technology, Seoul National University, Pyeongchang-gun, Gangwon-do, Korea; and Institute for Biomedical Sciences, Shinshu University, Kamiina, Nagano, Japan jaehan@snu.ac.kr.
authors.forename: Jae Yong
authors.initials: JY
authors.lastname: Han
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo; D002645:Chickens / Q000235:genetics*; D000066473:Embryonic Germ Cells; D005260:Female; D005786:Gene Expression Regulation; D018390:Gene Targeting; D016678:Genome; D049452:Green Fluorescent Proteins; D008297:Male; D030781:Organisms, Genetically Modified; D045522:Recombinases / Q000378:metabolism; D011995:Recombination, Genetic / Q000235:genetics*
chemical_list: D045522:Recombinases; D049452:Green Fluorescent Proteins
keywords: piggyBac transposition; primordial germ cell; transgene integration; transgenic chicken