title: Highly effective ex vivo gene manipulation to study kidney development using self-complementary adenoassociated viruses.
abstract: BACKGROUND
  Ex vivo culture of intact embryonic kidney has become a powerful system for studying renal development. However, few methods have been available for gene manipulation and have impeded the identification and investigation of genes in this developmental process.
  
  
  RESULTS
  Here we systemically compared eight different serotypes of pseudotyped self-complementary adenoassociated viruses (scAAVs) transduction in cultured embryonic kidney with a modified culture procedure. We demonstrated that scAAV was highly effective in delivering genes into and expressing in compacted tissues. scAAV serotypes 2 and 8 exhibited higher efficiency of transduction compared to others. Expression kinetics assay revealed that scAAV can be used for gene manipulation at the study of UB branching and nephrogenesis. Repressing WT1 in cultured kidney using shRNA impairs tubule formation. We for the first time employed and validated scAAV as a gene delivery tool in cultured kidney.
  
  
  CONCLUSIONS
  These findings are expected to expedite the use of the ex vivo embryonic kidney cultures for kidney development research. For other ex vivo cultured organ models, scAAV could also be a promising tool for organogenesis study.
authors.affiliation: Core Facility of Genetically Engineered Mice, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, China.
authors.forename: Tie-Lin
authors.initials: TL
authors.lastname: Chen
authors.affiliation: Core Facility of Genetically Engineered Mice, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, China.
authors.forename: Hong-Lian
authors.initials: HL
authors.lastname: Wang
authors.affiliation: Core Facility of Genetically Engineered Mice, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, China.
authors.forename: Yun-Hong
authors.initials: YH
authors.lastname: Liu
authors.affiliation: Core Facility of Genetically Engineered Mice, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, China.
authors.forename: Yin
authors.initials: Y
authors.lastname: Fang
authors.affiliation: Core Facility of Genetically Engineered Mice, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, China.
authors.forename: Rui-Zhi
authors.initials: RZ
authors.lastname: Tan
authors.affiliation: College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Pu-Hui
authors.initials: PH
authors.lastname: Zhou
authors.affiliation: Core Facility of Genetically Engineered Mice, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, China.
authors.forename: Qin
authors.initials: Q
authors.lastname: Zhou
authors.affiliation: Departments of Dermatology, West China Hospital, Sichuan University, Sichuan 610041, China.
authors.forename: Xiao-Yan
authors.initials: XY
authors.lastname: Lv
mesh_terms: D000818:Animals; D000229:Dependovirus / Q000235:genetics*; D057809:HEK293 Cells; D006801:Humans; D007668:Kidney / Q000254:growth & development / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D009924:Organ Culture Techniques / Q000379:methods; D014161:Transduction, Genetic / Q000379:methods*