Construction and expression verification of mouse spermatogenic cell-specific expression vector AAV-Dazl-RFP-Flag
Objective To study the effect of spermatogenic cell-specific promoter Dazl in Spermatogenic cells. Constructing the eukaryotic reporter gene expression vector AAV-Dazl-RFP-Flag containing spermatogenic cell-specific promoter Dazl. Methods The Adeno-associated virus vector AAV-CMV-RFP-Flag was modifi...
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| Format: | Article |
| Language: | zho |
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Institute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College.
2020-06-01
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| Series: | Jichu yixue yu linchuang |
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| Online Access: | http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/a200234.pdf |
| _version_ | 1797365756015411200 |
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| author | LI Ya-hui, LI Kai, LUO Yan-yun, LI Meng-zhen, MANG Xin-yu, SONG Wei, YANG Tao |
| author_facet | LI Ya-hui, LI Kai, LUO Yan-yun, LI Meng-zhen, MANG Xin-yu, SONG Wei, YANG Tao |
| author_sort | LI Ya-hui, LI Kai, LUO Yan-yun, LI Meng-zhen, MANG Xin-yu, SONG Wei, YANG Tao |
| collection | DOAJ |
| description | Objective To study the effect of spermatogenic cell-specific promoter Dazl in Spermatogenic cells. Constructing the eukaryotic reporter gene expression vector AAV-Dazl-RFP-Flag containing spermatogenic cell-specific promoter Dazl. Methods The Adeno-associated virus vector AAV-CMV-RFP-Flag was modified by homologous recombination to construct mouse permatogenic cell-specific expression vector AAV-Dazl-RFP-Flag. In vitro, GC1-spg wild-type cells and HEK-293T cells were transfected, and the expression specificity of AAV-Dazl-RFP-Flag in vitro was verified by immunofluorescence, q-PCR, and Western blot. In vivo, AAV-Dazl-RFP-Flag was packaged with adeno-associated virus (AAV), and AAV was injected into the testes of mouse through microinjection technology, then the expression specificity of AAV-Dazl-RFP-Flag in vivo was detected by q-PCR. Results Mouse Sper- matogenic cell-specific expression vector AAV-Dazl-RFP-Flag is specifically expressed in spermatogenic, cells thus provid providing a more efficient tool for gene therapy of male reproductive infertility. |
| first_indexed | 2024-03-08T16:53:28Z |
| format | Article |
| id | doaj.art-ef49924adb1545afb8bf40b1077f0e85 |
| institution | Directory Open Access Journal |
| issn | 1001-6325 |
| language | zho |
| last_indexed | 2024-03-08T16:53:28Z |
| publishDate | 2020-06-01 |
| publisher | Institute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College. |
| record_format | Article |
| series | Jichu yixue yu linchuang |
| spelling | doaj.art-ef49924adb1545afb8bf40b1077f0e852024-01-05T03:15:51ZzhoInstitute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College.Jichu yixue yu linchuang1001-63252020-06-01406753758Construction and expression verification of mouse spermatogenic cell-specific expression vector AAV-Dazl-RFP-FlagLI Ya-hui, LI Kai, LUO Yan-yun, LI Meng-zhen, MANG Xin-yu, SONG Wei, YANG Tao01. Department of Biochemistry and Molecular Biology, Shanxi Medical University, Taiyuan 030001;;2. National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences CAMS, School of Basic Medicine PUMC, Beijing 100005, ChinaObjective To study the effect of spermatogenic cell-specific promoter Dazl in Spermatogenic cells. Constructing the eukaryotic reporter gene expression vector AAV-Dazl-RFP-Flag containing spermatogenic cell-specific promoter Dazl. Methods The Adeno-associated virus vector AAV-CMV-RFP-Flag was modified by homologous recombination to construct mouse permatogenic cell-specific expression vector AAV-Dazl-RFP-Flag. In vitro, GC1-spg wild-type cells and HEK-293T cells were transfected, and the expression specificity of AAV-Dazl-RFP-Flag in vitro was verified by immunofluorescence, q-PCR, and Western blot. In vivo, AAV-Dazl-RFP-Flag was packaged with adeno-associated virus (AAV), and AAV was injected into the testes of mouse through microinjection technology, then the expression specificity of AAV-Dazl-RFP-Flag in vivo was detected by q-PCR. Results Mouse Sper- matogenic cell-specific expression vector AAV-Dazl-RFP-Flag is specifically expressed in spermatogenic, cells thus provid providing a more efficient tool for gene therapy of male reproductive infertility.http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/a200234.pdfdazl|promoter|gene therapy|spermatogenesis |
| spellingShingle | LI Ya-hui, LI Kai, LUO Yan-yun, LI Meng-zhen, MANG Xin-yu, SONG Wei, YANG Tao Construction and expression verification of mouse spermatogenic cell-specific expression vector AAV-Dazl-RFP-Flag Jichu yixue yu linchuang dazl|promoter|gene therapy|spermatogenesis |
| title | Construction and expression verification of mouse spermatogenic cell-specific expression vector AAV-Dazl-RFP-Flag |
| title_full | Construction and expression verification of mouse spermatogenic cell-specific expression vector AAV-Dazl-RFP-Flag |
| title_fullStr | Construction and expression verification of mouse spermatogenic cell-specific expression vector AAV-Dazl-RFP-Flag |
| title_full_unstemmed | Construction and expression verification of mouse spermatogenic cell-specific expression vector AAV-Dazl-RFP-Flag |
| title_short | Construction and expression verification of mouse spermatogenic cell-specific expression vector AAV-Dazl-RFP-Flag |
| title_sort | construction and expression verification of mouse spermatogenic cell specific expression vector aav dazl rfp flag |
| topic | dazl|promoter|gene therapy|spermatogenesis |
| url | http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/a200234.pdf |
| work_keys_str_mv | AT liyahuilikailuoyanyunlimengzhenmangxinyusongweiyangtao constructionandexpressionverificationofmousespermatogeniccellspecificexpressionvectoraavdazlrfpflag |