Transgenic mice Cre-dependently expressing mutant polymerase-gamma: novel test-system for pharmacological study of mitoprotective drugs
Introduction: PolG-alpha is a nuclear-encoded enzyme which provides replication and repair of mitochondrial DNA. D257A mutation of PolG-alpha leads to change in the N-terminal ”proofreading” domain, which deprives the enzyme of 3′-5′ exonuclease activity, resulting in accumulation of mutations in th...
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Belgorod National Research University
2021-09-01
|
| Series: | Research Results in Pharmacology |
| Online Access: | https://rrpharmacology.pensoft.net/article/72784/download/pdf/ |
| _version_ | 1827615928503762944 |
|---|---|
| author | Marina V. Kubekina Yulia Yu. Silaeva Alexandra V. Bruter Diana S. Korshunova Leonid A. Ilchuk Yulia D. Okulova Mariya O. Soldatova Evgeniya Seryogina Inga M. Kolesnik Polina A. Ukolova Mikhail V. Korokin Alexey V. Deykin |
| author_facet | Marina V. Kubekina Yulia Yu. Silaeva Alexandra V. Bruter Diana S. Korshunova Leonid A. Ilchuk Yulia D. Okulova Mariya O. Soldatova Evgeniya Seryogina Inga M. Kolesnik Polina A. Ukolova Mikhail V. Korokin Alexey V. Deykin |
| author_sort | Marina V. Kubekina |
| collection | DOAJ |
| description | Introduction: PolG-alpha is a nuclear-encoded enzyme which provides replication and repair of mitochondrial DNA. D257A mutation of PolG-alpha leads to change in the N-terminal ”proofreading” domain, which deprives the enzyme of 3′-5′ exonuclease activity, resulting in accumulation of mutations in the mitochondrial genome. Materials and methods: Murine zygotes were microinjected with transgene construction carrying mutant murine Polg coding sequence and GFP coding sequence by a loxP-flanked STOP-cassette. Two Cre-activator strains, CMV-Cre (systemic activation) and Tie2-Cre (endothelial activation), were used for activation of the transgene. To confirm the insertion and Cre-dependent activation of the transgene, genotyping and qPCR copy number measurement of mutant Polg were performed, and GFP fluorescence was assessed. Results: Two primary transgenic animals were used as the founders for two lines with copy numbers of transgene ~7 and ~5. After systemic activation, the number of the transgene copies decreases to ~1.0 while endothelial specific activation does not affect the number of transgene copies in tail tissue. Discussion: A murine model with spatial control of mutant Polg expression has been developed. To our knowledge, this is the first transgenic model of tissue-specific mitochondrial dysfunction. Conclusion: Transgenic mice Cre-dependent expressing mutant polymerase-gamma are a novel test-system for studying mitochondrial biology and efficacy of mitoprotective drugs. |
| first_indexed | 2024-03-09T09:18:37Z |
| format | Article |
| id | doaj.art-2f3fb4bdef8948059ade8b9c4c2824c0 |
| institution | Directory Open Access Journal |
| issn | 2658-381X |
| language | English |
| last_indexed | 2024-03-09T09:18:37Z |
| publishDate | 2021-09-01 |
| publisher | Belgorod National Research University |
| record_format | Article |
| series | Research Results in Pharmacology |
| spelling | doaj.art-2f3fb4bdef8948059ade8b9c4c2824c02023-12-02T07:03:19ZengBelgorod National Research UniversityResearch Results in Pharmacology2658-381X2021-09-0173333910.3897/rrpharmacology.7.7278472784Transgenic mice Cre-dependently expressing mutant polymerase-gamma: novel test-system for pharmacological study of mitoprotective drugsMarina V. Kubekina0Yulia Yu. Silaeva1Alexandra V. Bruter2Diana S. Korshunova3Leonid A. Ilchuk4Yulia D. Okulova5Mariya O. Soldatova6Evgeniya Seryogina7Inga M. Kolesnik8Polina A. Ukolova9Mikhail V. Korokin10Alexey V. Deykin11Institute of Gene Biology, Russian Academy of SciencesInstitute of Gene Biology, Russian Academy of SciencesInstitute of Gene Biology, Russian Academy of SciencesInstitute of Gene Biology, Russian Academy of SciencesInstitute of Gene Biology, Russian Academy of SciencesInstitute of Gene Biology, Russian Academy of SciencesKursk State Medical UniversityOrel State UniversityBelgorod State National Research UniversityOrel State UniversityBelgorod State National Research UniversityInstitute of Gene Biology, Russian Academy of SciencesIntroduction: PolG-alpha is a nuclear-encoded enzyme which provides replication and repair of mitochondrial DNA. D257A mutation of PolG-alpha leads to change in the N-terminal ”proofreading” domain, which deprives the enzyme of 3′-5′ exonuclease activity, resulting in accumulation of mutations in the mitochondrial genome. Materials and methods: Murine zygotes were microinjected with transgene construction carrying mutant murine Polg coding sequence and GFP coding sequence by a loxP-flanked STOP-cassette. Two Cre-activator strains, CMV-Cre (systemic activation) and Tie2-Cre (endothelial activation), were used for activation of the transgene. To confirm the insertion and Cre-dependent activation of the transgene, genotyping and qPCR copy number measurement of mutant Polg were performed, and GFP fluorescence was assessed. Results: Two primary transgenic animals were used as the founders for two lines with copy numbers of transgene ~7 and ~5. After systemic activation, the number of the transgene copies decreases to ~1.0 while endothelial specific activation does not affect the number of transgene copies in tail tissue. Discussion: A murine model with spatial control of mutant Polg expression has been developed. To our knowledge, this is the first transgenic model of tissue-specific mitochondrial dysfunction. Conclusion: Transgenic mice Cre-dependent expressing mutant polymerase-gamma are a novel test-system for studying mitochondrial biology and efficacy of mitoprotective drugs.https://rrpharmacology.pensoft.net/article/72784/download/pdf/ |
| spellingShingle | Marina V. Kubekina Yulia Yu. Silaeva Alexandra V. Bruter Diana S. Korshunova Leonid A. Ilchuk Yulia D. Okulova Mariya O. Soldatova Evgeniya Seryogina Inga M. Kolesnik Polina A. Ukolova Mikhail V. Korokin Alexey V. Deykin Transgenic mice Cre-dependently expressing mutant polymerase-gamma: novel test-system for pharmacological study of mitoprotective drugs Research Results in Pharmacology |
| title | Transgenic mice Cre-dependently expressing mutant polymerase-gamma: novel test-system for pharmacological study of mitoprotective drugs |
| title_full | Transgenic mice Cre-dependently expressing mutant polymerase-gamma: novel test-system for pharmacological study of mitoprotective drugs |
| title_fullStr | Transgenic mice Cre-dependently expressing mutant polymerase-gamma: novel test-system for pharmacological study of mitoprotective drugs |
| title_full_unstemmed | Transgenic mice Cre-dependently expressing mutant polymerase-gamma: novel test-system for pharmacological study of mitoprotective drugs |
| title_short | Transgenic mice Cre-dependently expressing mutant polymerase-gamma: novel test-system for pharmacological study of mitoprotective drugs |
| title_sort | transgenic mice cre dependently expressing mutant polymerase gamma novel test system for pharmacological study of mitoprotective drugs |
| url | https://rrpharmacology.pensoft.net/article/72784/download/pdf/ |
| work_keys_str_mv | AT marinavkubekina transgenicmicecredependentlyexpressingmutantpolymerasegammanoveltestsystemforpharmacologicalstudyofmitoprotectivedrugs AT yuliayusilaeva transgenicmicecredependentlyexpressingmutantpolymerasegammanoveltestsystemforpharmacologicalstudyofmitoprotectivedrugs AT alexandravbruter transgenicmicecredependentlyexpressingmutantpolymerasegammanoveltestsystemforpharmacologicalstudyofmitoprotectivedrugs AT dianaskorshunova transgenicmicecredependentlyexpressingmutantpolymerasegammanoveltestsystemforpharmacologicalstudyofmitoprotectivedrugs AT leonidailchuk transgenicmicecredependentlyexpressingmutantpolymerasegammanoveltestsystemforpharmacologicalstudyofmitoprotectivedrugs AT yuliadokulova transgenicmicecredependentlyexpressingmutantpolymerasegammanoveltestsystemforpharmacologicalstudyofmitoprotectivedrugs AT mariyaosoldatova transgenicmicecredependentlyexpressingmutantpolymerasegammanoveltestsystemforpharmacologicalstudyofmitoprotectivedrugs AT evgeniyaseryogina transgenicmicecredependentlyexpressingmutantpolymerasegammanoveltestsystemforpharmacologicalstudyofmitoprotectivedrugs AT ingamkolesnik transgenicmicecredependentlyexpressingmutantpolymerasegammanoveltestsystemforpharmacologicalstudyofmitoprotectivedrugs AT polinaaukolova transgenicmicecredependentlyexpressingmutantpolymerasegammanoveltestsystemforpharmacologicalstudyofmitoprotectivedrugs AT mikhailvkorokin transgenicmicecredependentlyexpressingmutantpolymerasegammanoveltestsystemforpharmacologicalstudyofmitoprotectivedrugs AT alexeyvdeykin transgenicmicecredependentlyexpressingmutantpolymerasegammanoveltestsystemforpharmacologicalstudyofmitoprotectivedrugs |