Highly Effective Ex Vivo Gene Manipulation to Study Kidney Development Using Self-Complementary Adenoassociated Viruses
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 systemical...
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| Format: | Article |
| Language: | English |
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Wiley
2014-01-01
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| Series: | The Scientific World Journal |
| Online Access: | http://dx.doi.org/10.1155/2014/682189 |
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| author | Tie-Lin Chen Hong-Lian Wang Yun-Hong Liu Yin Fang Rui-Zhi Tan Pu-Hui Zhou Qin Zhou Xiao-Yan Lv |
| author_facet | Tie-Lin Chen Hong-Lian Wang Yun-Hong Liu Yin Fang Rui-Zhi Tan Pu-Hui Zhou Qin Zhou Xiao-Yan Lv |
| author_sort | Tie-Lin Chen |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-b4a2ea876c8e4a85a84e0ae42ef46381 |
| institution | Kabale University |
| issn | 2356-6140 1537-744X |
| language | English |
| publishDate | 2014-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | The Scientific World Journal |
| spelling | doaj-art-b4a2ea876c8e4a85a84e0ae42ef463812025-02-03T01:23:03ZengWileyThe Scientific World Journal2356-61401537-744X2014-01-01201410.1155/2014/682189682189Highly Effective Ex Vivo Gene Manipulation to Study Kidney Development Using Self-Complementary Adenoassociated VirusesTie-Lin Chen0Hong-Lian Wang1Yun-Hong Liu2Yin Fang3Rui-Zhi Tan4Pu-Hui Zhou5Qin Zhou6Xiao-Yan Lv7Core Facility of Genetically Engineered Mice, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, ChinaCore Facility of Genetically Engineered Mice, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, ChinaCore Facility of Genetically Engineered Mice, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, ChinaCore Facility of Genetically Engineered Mice, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, ChinaCore Facility of Genetically Engineered Mice, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, ChinaCollege of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, ChinaCore Facility of Genetically Engineered Mice, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, ChinaDepartments of Dermatology, West China Hospital, Sichuan University, Sichuan 610041, ChinaBackground. 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.http://dx.doi.org/10.1155/2014/682189 |
| spellingShingle | Tie-Lin Chen Hong-Lian Wang Yun-Hong Liu Yin Fang Rui-Zhi Tan Pu-Hui Zhou Qin Zhou Xiao-Yan Lv Highly Effective Ex Vivo Gene Manipulation to Study Kidney Development Using Self-Complementary Adenoassociated Viruses The Scientific World Journal |
| title | Highly Effective Ex Vivo Gene Manipulation to Study Kidney Development Using Self-Complementary Adenoassociated Viruses |
| title_full | Highly Effective Ex Vivo Gene Manipulation to Study Kidney Development Using Self-Complementary Adenoassociated Viruses |
| title_fullStr | Highly Effective Ex Vivo Gene Manipulation to Study Kidney Development Using Self-Complementary Adenoassociated Viruses |
| title_full_unstemmed | Highly Effective Ex Vivo Gene Manipulation to Study Kidney Development Using Self-Complementary Adenoassociated Viruses |
| title_short | Highly Effective Ex Vivo Gene Manipulation to Study Kidney Development Using Self-Complementary Adenoassociated Viruses |
| title_sort | highly effective ex vivo gene manipulation to study kidney development using self complementary adenoassociated viruses |
| url | http://dx.doi.org/10.1155/2014/682189 |
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