title: Efficient Generation of Mice with Consistent Transgene Expression by FEEST.
abstract: Transgenic mouse models are widely used in biomedical research; however, current techniques for producing transgenic mice are limited due to the unpredictable nature of transgene expression. Here, we report a novel, highly efficient technique for the generation of transgenic mice with single-copy integration of the transgene and guaranteed expression of the gene-of-interest (GOI). We refer to this technique as functionally enriched ES cell transgenics, or FEEST. ES cells harboring an inducible Cre gene enabled the efficient selection of transgenic ES cell clones using hygromycin before Cre-mediated recombination. Expression of the GOI was confirmed by assaying for the GFP after Cre recombination. As a proof-of-principle, we produced a transgenic mouse line containing Cre-activatable tTA (cl-tTA6). This tTA mouse model was able to induce tumor formation when crossed with a transgenic mouse line containing a doxycycline-inducible oncogene. We also showed that the cl-tTA6 mouse is a valuable tool for faithfully recapitulating the clinical course of tumor development. We showed that FEEST can be easily adapted for other genes by preparing a transgenic mouse model of conditionally activatable EGFR L858R. Thus, FEEST is a technique with the potential to generate transgenic mouse models at a genome-wide scale.
authors.affiliation: College of Life Sciences, Beijing Normal University, Beijing, 100875, China.|National Institute of Biological Sciences, Beijing, Beijing 102206, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Gao
authors.affiliation: National Institute of Biological Sciences, Beijing, Beijing 102206, China.
authors.forename: Yonghua
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: School of Life Science, Tsinghua University, Beijing 100084, China.
authors.forename: Libing
authors.initials: L
authors.lastname: Mu
authors.affiliation: National Institute of Biological Sciences, Collaborative Innovation Center for Cancer Medicine, Beijing, 102206, China.
authors.forename: Yanbin
authors.initials: Y
authors.lastname: Liu
authors.affiliation: National Institute of Biological Sciences, Beijing, Beijing 102206, China.
authors.forename: Fengchao
authors.initials: F
authors.lastname: Wang
authors.affiliation: Beijing Ditan Hospital, Beijing 100015, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Wang
authors.affiliation: General Hospital of People's Liberation Army, Beijing 100853, China.
authors.forename: Aiqun
authors.initials: A
authors.lastname: Zhang
authors.affiliation: National Institute of Biological Sciences, Beijing, Beijing 102206, China.
authors.forename: Nan
authors.initials: N
authors.lastname: Tang
authors.affiliation: National Institute of Biological Sciences, Beijing, Beijing 102206, China.
authors.forename: Ting
authors.initials: T
authors.lastname: Chen
authors.affiliation: National Institute of Biological Sciences, Beijing, Beijing 102206, China.
authors.forename: Minmin
authors.initials: M
authors.lastname: Luo
authors.affiliation: Beijing Tongren Hospital, Capital Medical University, Beijing 100730, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Yu
authors.affiliation: Tongji University, Shanghai, 200092, China.
authors.forename: Shaorong
authors.initials: S
authors.lastname: Gao
authors.affiliation: National Institute of Biological Sciences, Beijing, Beijing 102206, China.|National Institute of Biological Sciences, Collaborative Innovation Center for Cancer Medicine, Beijing, 102206, China.
authors.forename: Liang
authors.initials: L
authors.lastname: Chen
mesh_terms: D000818:Animals; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D066246:ErbB Receptors / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008822:Mice, Transgenic; D023421:Models, Animal; D011995:Recombination, Genetic; D019076:Transgenes / Q000235:genetics*; D018631:ras Proteins / Q000235:genetics
chemical_list: C541485:EGFR protein, mouse; D066246:ErbB Receptors; D018631:ras Proteins