Production of Lentiviral Vector Expressing MicroRNA-148b
Background: Micro (mi)RNAs are non-coding endogenous RNAs which regulate gene expression by hybridization to specific binding sites in target mRNA sequences. Since several miRNAs are involved in proliferation and differentiation, miRNA-based therapies could be promising approach in regenerative medi...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | fas |
| Published: |
Isfahan University of Medical Sciences
2017-08-01
|
| Series: | مجله دانشکده پزشکی اصفهان |
| Subjects: | |
| Online Access: | http://jims.mui.ac.ir/index.php/jims/article/view/7895 |
| _version_ | 1797764795200438272 |
|---|---|
| author | Samaneh Mollazadeh Vajiheh Neshati Bibi Sedigheh Fazly Bazzaz Majid Mojarrad Mohammad Amin Kerachian |
| author_facet | Samaneh Mollazadeh Vajiheh Neshati Bibi Sedigheh Fazly Bazzaz Majid Mojarrad Mohammad Amin Kerachian |
| author_sort | Samaneh Mollazadeh |
| collection | DOAJ |
| description | Background: Micro (mi)RNAs are non-coding endogenous RNAs which regulate gene expression by hybridization to specific binding sites in target mRNA sequences. Since several miRNAs are involved in proliferation and differentiation, miRNA-based therapies could be promising approach in regenerative medicine. Among different vehicles, lentiviral vector system is suitable for miRNA delivery. Besides, it is shown that miRNA-148b is involved in osteogenic differentiation. In this study, designing and cloning of miR-148b to lentiviral vector were investigated.
Methods: We introduced miRNA-148b-3p/-5p into lentiviral vector through cloning producers. The sequences of lentiviral vectors carrying miRNA-148b were checked via analytical digestion as well as Sanger DNA sequencing. In the following, produced lentiviral vectors were used for mesenchymal stem cells transduction.
Findings: Designed miR-148b-3p/-5p successfully cloned to the shuttle. Correctness and absence of any unintended mutations of lentiviral shuttle carrying miRNA-148b3p/-5p were confirmed followed by lentiviral production. Expression of enhanced green fluorescent protein (eGFP) demonstrated high efficiency of transfection as well as transduction.
Conclusion: Viral vectors constructed in this study could be used for investigation of osteogenesis. |
| first_indexed | 2024-03-12T20:00:50Z |
| format | Article |
| id | doaj.art-6a1c2462808e442983a253c8c49c92c4 |
| institution | Directory Open Access Journal |
| issn | 1027-7595 1735-854X |
| language | fas |
| last_indexed | 2024-03-12T20:00:50Z |
| publishDate | 2017-08-01 |
| publisher | Isfahan University of Medical Sciences |
| record_format | Article |
| series | مجله دانشکده پزشکی اصفهان |
| spelling | doaj.art-6a1c2462808e442983a253c8c49c92c42023-08-02T02:26:01ZfasIsfahan University of Medical Sciencesمجله دانشکده پزشکی اصفهان1027-75951735-854X2017-08-01354347017062563Production of Lentiviral Vector Expressing MicroRNA-148bSamaneh Mollazadeh0Vajiheh Neshati1Bibi Sedigheh Fazly Bazzaz2Majid Mojarrad3Mohammad Amin Kerachian4PhD Candidate, Biotechnology Research Center, Mashhad University of Medical Sciences, Mashhad, IranPhD Candidate, Biotechnology Research Center, Mashhad University of Medical Sciences, Mashhad, IranProfessor, Biotechnology Research Center, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, IranAssistant Professor, Medical Genetics Research Center AND Department of Medical Genetics, School of Medicine, Mashhad University of Medical Sciences, Mashhad, IranAssistant Professor, Medical Genetics Research Center AND Department of Medical Genetics, School of Medicine, Mashhad University of Medical Sciences, Mashhad, IranBackground: Micro (mi)RNAs are non-coding endogenous RNAs which regulate gene expression by hybridization to specific binding sites in target mRNA sequences. Since several miRNAs are involved in proliferation and differentiation, miRNA-based therapies could be promising approach in regenerative medicine. Among different vehicles, lentiviral vector system is suitable for miRNA delivery. Besides, it is shown that miRNA-148b is involved in osteogenic differentiation. In this study, designing and cloning of miR-148b to lentiviral vector were investigated. Methods: We introduced miRNA-148b-3p/-5p into lentiviral vector through cloning producers. The sequences of lentiviral vectors carrying miRNA-148b were checked via analytical digestion as well as Sanger DNA sequencing. In the following, produced lentiviral vectors were used for mesenchymal stem cells transduction. Findings: Designed miR-148b-3p/-5p successfully cloned to the shuttle. Correctness and absence of any unintended mutations of lentiviral shuttle carrying miRNA-148b3p/-5p were confirmed followed by lentiviral production. Expression of enhanced green fluorescent protein (eGFP) demonstrated high efficiency of transfection as well as transduction. Conclusion: Viral vectors constructed in this study could be used for investigation of osteogenesis.http://jims.mui.ac.ir/index.php/jims/article/view/7895CloningMicoRNAMiR-148bShuttle vectorsLentivirus |
| spellingShingle | Samaneh Mollazadeh Vajiheh Neshati Bibi Sedigheh Fazly Bazzaz Majid Mojarrad Mohammad Amin Kerachian Production of Lentiviral Vector Expressing MicroRNA-148b مجله دانشکده پزشکی اصفهان Cloning MicoRNA MiR-148b Shuttle vectors Lentivirus |
| title | Production of Lentiviral Vector Expressing MicroRNA-148b |
| title_full | Production of Lentiviral Vector Expressing MicroRNA-148b |
| title_fullStr | Production of Lentiviral Vector Expressing MicroRNA-148b |
| title_full_unstemmed | Production of Lentiviral Vector Expressing MicroRNA-148b |
| title_short | Production of Lentiviral Vector Expressing MicroRNA-148b |
| title_sort | production of lentiviral vector expressing microrna 148b |
| topic | Cloning MicoRNA MiR-148b Shuttle vectors Lentivirus |
| url | http://jims.mui.ac.ir/index.php/jims/article/view/7895 |
| work_keys_str_mv | AT samanehmollazadeh productionoflentiviralvectorexpressingmicrorna148b AT vajihehneshati productionoflentiviralvectorexpressingmicrorna148b AT bibisedighehfazlybazzaz productionoflentiviralvectorexpressingmicrorna148b AT majidmojarrad productionoflentiviralvectorexpressingmicrorna148b AT mohammadaminkerachian productionoflentiviralvectorexpressingmicrorna148b |