title: A modified TALEN-based strategy for rapidly and efficiently generating knockout mice for kidney development studies.
abstract: The transcription activator-like effector nucleases (TALENs) strategy has been widely used to delete and mutate genes in vitro. This strategy has begun to be used for in vivo systemic gene manipulation, but not in an organ-specific manner. In this study, we developed a modified, highly efficient TALEN strategy using a dual-fluorescence reporter. We used this modified strategy and, within 5 weeks, we successfully generated kidney proximal tubule-specific gene Ttc36 homozygous knockout mice. Unilateral nephrectomy was performed on the 6-week-old founders (F0) to identify the knockout genotype prior to the birth of the offspring. This strategy was found to have little effect on reproduction in the knockout mice and inheritability of the knockout genotypes. The modified TALEN knockout strategy in combination with unilateral nephrectomy can be readily used for studies of gene function in kidney development and diseases.
authors.affiliation: Core Facility of Genetically Engineered Mice, Regenerative Medicine Research Center, West China Hospital, West China Medical School, Sichuan University, Chengdu, Sichuan, China.
authors.forename: Yunhong
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Dermatology, West China Hospital, West China Medical School, Sichuan University, Chengdu, Sichuan, China.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Lv
authors.affiliation: Core Facility of Genetically Engineered Mice, Regenerative Medicine Research Center, West China Hospital, West China Medical School, Sichuan University, Chengdu, Sichuan, China.
authors.forename: Ruizhi
authors.initials: R
authors.lastname: Tan
authors.affiliation: Lab of Molecular Nephrology, Chongqing Medical University, Chongqing, China.
authors.forename: Tianming
authors.initials: T
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, Sichuan University, Chengdu, Sichuan, China.
authors.forename: Tielin
authors.initials: T
authors.lastname: Chen
authors.affiliation: Core Facility of Genetically Engineered Mice, Regenerative Medicine Research Center, West China Hospital, West China Medical School, Sichuan University, Chengdu, Sichuan, China.
authors.forename: Mi
authors.initials: M
authors.lastname: Li
authors.affiliation: Core Facility of Genetically Engineered Mice, Regenerative Medicine Research Center, West China Hospital, West China Medical School, Sichuan University, Chengdu, Sichuan, China.
authors.forename: Yuhang
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Lab of Molecular Nephrology, Chongqing Medical University, Chongqing, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Nie
authors.affiliation: Lab of Molecular Nephrology, Chongqing Medical University, Chongqing, China.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Wang
authors.affiliation: Lab of Molecular Nephrology, Chongqing Medical University, Chongqing, China.
authors.forename: Puhui
authors.initials: P
authors.lastname: Zhou
authors.affiliation: Core Facility of Genetically Engineered Mice, Regenerative Medicine Research Center, West China Hospital, West China Medical School, Sichuan University, Chengdu, Sichuan, China.
authors.forename: Mianzhi
authors.initials: M
authors.lastname: Chen
authors.affiliation: Core Facility of Genetically Engineered Mice, Regenerative Medicine Research Center, West China Hospital, West China Medical School, Sichuan University, Chengdu, Sichuan, China.
authors.forename: Qin
authors.initials: Q
authors.lastname: Zhou
mesh_terms: D000818:Animals; D001483:Base Sequence; D002467:Cell Nucleus / Q000235:genetics; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D016368:Frameshift Mutation; D055786:Gene Knockout Techniques / Q000379:methods*; D017930:Genes, Reporter / Q000235:genetics; D057809:HEK293 Cells; D059765:Homologous Recombination / Q000235:genetics; D006720:Homozygote; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000254:growth & development* / Q000378:metabolism / Q000601:surgery; D051379:Mice; D009392:Nephrectomy; D010641:Phenotype; D013997:Time Factors
chemical_list: D004262:DNA Restriction Enzymes