Microinjection of antisense oligonucleotides into living mouse testis enables lncRNA function study
Abstract Background Long non-coding RNAs (lncRNAs) have been the focus of ongoing research in a diversity of cellular processes. LncRNAs are abundant in mammalian testis, but their biological function remains poorly known. Results Here, we established an antisense oligonucleotides (ASOs)-based targe...
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| Format: | Article |
| Language: | English |
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BMC
2021-12-01
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| Series: | Cell & Bioscience |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13578-021-00717-y |
| _version_ | 1819177223868907520 |
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| author | Zhaohui Chen Li Ling Xiaolian Shi Wu Li Huicong Zhai Zhenlong Kang Bangjin Zheng Jiaqi Zhu Suni Ye Hao Wang Lingxiu Tong Juan Ni Chaoyang Huang Yang Li Ke Zheng |
| author_facet | Zhaohui Chen Li Ling Xiaolian Shi Wu Li Huicong Zhai Zhenlong Kang Bangjin Zheng Jiaqi Zhu Suni Ye Hao Wang Lingxiu Tong Juan Ni Chaoyang Huang Yang Li Ke Zheng |
| author_sort | Zhaohui Chen |
| collection | DOAJ |
| description | Abstract Background Long non-coding RNAs (lncRNAs) have been the focus of ongoing research in a diversity of cellular processes. LncRNAs are abundant in mammalian testis, but their biological function remains poorly known. Results Here, we established an antisense oligonucleotides (ASOs)-based targeting approach that can efficiently knock down lncRNA in living mouse testis. We cloned the full-length transcript of lncRNA Tsx (testis-specific X-linked) and defined its testicular localization pattern. Microinjection of ASOs through seminiferous tubules in vivo significantly lowered the Tsx levels in both nucleus and cytoplasm. This effect lasted no less than 10 days, conducive to the generation and maintenance of phenotype. Importantly, ASOs performed better in depleting the nuclear Tsx and sustained longer effect than small interfering RNAs (siRNAs). In addition to the observation of an elevated number of apoptotic germ cells upon ASOs injection, which recapitulates the documented description of Tsx knockout, we also found a specific loss of meiotic spermatocytes despite overall no impact on meiosis and male fertility. Conclusions Our study detailed the characterization of Tsx and illustrates ASOs as an advantageous tool to functionally interrogate lncRNAs in spermatogenesis. |
| first_indexed | 2024-12-22T21:23:15Z |
| format | Article |
| id | doaj.art-9edeaefaa9b543a58b85ef6b73bc7477 |
| institution | Directory Open Access Journal |
| issn | 2045-3701 |
| language | English |
| last_indexed | 2024-12-22T21:23:15Z |
| publishDate | 2021-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Cell & Bioscience |
| spelling | doaj.art-9edeaefaa9b543a58b85ef6b73bc74772022-12-21T18:12:08ZengBMCCell & Bioscience2045-37012021-12-0111111110.1186/s13578-021-00717-yMicroinjection of antisense oligonucleotides into living mouse testis enables lncRNA function studyZhaohui Chen0Li Ling1Xiaolian Shi2Wu Li3Huicong Zhai4Zhenlong Kang5Bangjin Zheng6Jiaqi Zhu7Suni Ye8Hao Wang9Lingxiu Tong10Juan Ni11Chaoyang Huang12Yang Li13Ke Zheng14State Key Laboratory of Reproductive Medicine, Nanjing Medical UniversityState Key Laboratory of Reproductive Medicine, Nanjing Medical UniversityState Key Laboratory of Reproductive Medicine, Nanjing Medical UniversityState Key Laboratory of Reproductive Medicine, Nanjing Medical UniversityState Key Laboratory of Reproductive Medicine, Nanjing Medical UniversityState Key Laboratory of Reproductive Medicine, Nanjing Medical UniversityState Key Laboratory of Reproductive Medicine, Nanjing Medical UniversityState Key Laboratory of Reproductive Medicine, Nanjing Medical UniversityState Key Laboratory of Reproductive Medicine, Nanjing Medical UniversityState Key Laboratory of Reproductive Medicine, Nanjing Medical UniversityState Key Laboratory of Reproductive Medicine, Nanjing Medical UniversityDepartment of Obstetrics and Gynecology, the Affiliated Hospital of Hangzhou Normal UniversityDepartment of Cardiology, the First Affiliated Hospital, Zhejiang University School of MedicineState Key Laboratory of Reproductive Medicine, Nanjing Medical UniversityState Key Laboratory of Reproductive Medicine, Nanjing Medical UniversityAbstract Background Long non-coding RNAs (lncRNAs) have been the focus of ongoing research in a diversity of cellular processes. LncRNAs are abundant in mammalian testis, but their biological function remains poorly known. Results Here, we established an antisense oligonucleotides (ASOs)-based targeting approach that can efficiently knock down lncRNA in living mouse testis. We cloned the full-length transcript of lncRNA Tsx (testis-specific X-linked) and defined its testicular localization pattern. Microinjection of ASOs through seminiferous tubules in vivo significantly lowered the Tsx levels in both nucleus and cytoplasm. This effect lasted no less than 10 days, conducive to the generation and maintenance of phenotype. Importantly, ASOs performed better in depleting the nuclear Tsx and sustained longer effect than small interfering RNAs (siRNAs). In addition to the observation of an elevated number of apoptotic germ cells upon ASOs injection, which recapitulates the documented description of Tsx knockout, we also found a specific loss of meiotic spermatocytes despite overall no impact on meiosis and male fertility. Conclusions Our study detailed the characterization of Tsx and illustrates ASOs as an advantageous tool to functionally interrogate lncRNAs in spermatogenesis.https://doi.org/10.1186/s13578-021-00717-ylncRNA (long non-coding RNA)TsxASO (antisense oligonucleotide)Knock downTestis injection |
| spellingShingle | Zhaohui Chen Li Ling Xiaolian Shi Wu Li Huicong Zhai Zhenlong Kang Bangjin Zheng Jiaqi Zhu Suni Ye Hao Wang Lingxiu Tong Juan Ni Chaoyang Huang Yang Li Ke Zheng Microinjection of antisense oligonucleotides into living mouse testis enables lncRNA function study Cell & Bioscience lncRNA (long non-coding RNA) Tsx ASO (antisense oligonucleotide) Knock down Testis injection |
| title | Microinjection of antisense oligonucleotides into living mouse testis enables lncRNA function study |
| title_full | Microinjection of antisense oligonucleotides into living mouse testis enables lncRNA function study |
| title_fullStr | Microinjection of antisense oligonucleotides into living mouse testis enables lncRNA function study |
| title_full_unstemmed | Microinjection of antisense oligonucleotides into living mouse testis enables lncRNA function study |
| title_short | Microinjection of antisense oligonucleotides into living mouse testis enables lncRNA function study |
| title_sort | microinjection of antisense oligonucleotides into living mouse testis enables lncrna function study |
| topic | lncRNA (long non-coding RNA) Tsx ASO (antisense oligonucleotide) Knock down Testis injection |
| url | https://doi.org/10.1186/s13578-021-00717-y |
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