Cargando…
A Method for Assessing the Efficiency of the Nucleotide Excision Repair System Ex Vivo
The nucleotide excision repair (NER) is one of the main repair systems present in the cells of living organisms. It is responsible for the removal of a wide range of bulky DNA lesions. We succeeded in developing a method for assessing the efficiency of NER in the cell (ex vivo), which is a method ba...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
A.I. Gordeyev
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8526188/ https://www.ncbi.nlm.nih.gov/pubmed/34707905 http://dx.doi.org/10.32607/actanaturae.11430 |
_version_ | 1784585828082647040 |
---|---|
author | Popov, A. A. Orishchenko, K. E. Naumenko, K. N. Evdokimov, A. N. Petruseva, I. O. Lavrik, O. I. |
author_facet | Popov, A. A. Orishchenko, K. E. Naumenko, K. N. Evdokimov, A. N. Petruseva, I. O. Lavrik, O. I. |
author_sort | Popov, A. A. |
collection | PubMed |
description | The nucleotide excision repair (NER) is one of the main repair systems present in the cells of living organisms. It is responsible for the removal of a wide range of bulky DNA lesions. We succeeded in developing a method for assessing the efficiency of NER in the cell (ex vivo), which is a method based on the recovery of TagRFP fluorescent protein production through repair of the damage that blocks the expression of the appropriate gene. Our constructed plasmids containing bulky nFlu or nAnt lesions near the tagrfp gene promoter were shown to undergo repair in eukaryotic cells (HEK 293T) and that they can be used to analyze the efficiency of NER ex vivo. A comparative analysis of the time dependence of fluorescent cells accumulation after transfection with nFlu- and nAnt-DNA revealed that there are differences in how efficient their repair by the NER system of HEK 293T cells can be. The method can be used to assess the cell repair status and the repair efficiency of different structural damages. |
format | Online Article Text |
id | pubmed-8526188 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | A.I. Gordeyev |
record_format | MEDLINE/PubMed |
spelling | pubmed-85261882021-10-26 A Method for Assessing the Efficiency of the Nucleotide Excision Repair System Ex Vivo Popov, A. A. Orishchenko, K. E. Naumenko, K. N. Evdokimov, A. N. Petruseva, I. O. Lavrik, O. I. Acta Naturae Research Article The nucleotide excision repair (NER) is one of the main repair systems present in the cells of living organisms. It is responsible for the removal of a wide range of bulky DNA lesions. We succeeded in developing a method for assessing the efficiency of NER in the cell (ex vivo), which is a method based on the recovery of TagRFP fluorescent protein production through repair of the damage that blocks the expression of the appropriate gene. Our constructed plasmids containing bulky nFlu or nAnt lesions near the tagrfp gene promoter were shown to undergo repair in eukaryotic cells (HEK 293T) and that they can be used to analyze the efficiency of NER ex vivo. A comparative analysis of the time dependence of fluorescent cells accumulation after transfection with nFlu- and nAnt-DNA revealed that there are differences in how efficient their repair by the NER system of HEK 293T cells can be. The method can be used to assess the cell repair status and the repair efficiency of different structural damages. A.I. Gordeyev 2021 /pmc/articles/PMC8526188/ /pubmed/34707905 http://dx.doi.org/10.32607/actanaturae.11430 Text en Copyright ® 2021 National Research University Higher School of Economics. https://creativecommons.org/licenses/by/2.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Popov, A. A. Orishchenko, K. E. Naumenko, K. N. Evdokimov, A. N. Petruseva, I. O. Lavrik, O. I. A Method for Assessing the Efficiency of the Nucleotide Excision Repair System Ex Vivo |
title | A Method for Assessing the Efficiency of the Nucleotide Excision Repair System Ex Vivo |
title_full | A Method for Assessing the Efficiency of the Nucleotide Excision Repair System Ex Vivo |
title_fullStr | A Method for Assessing the Efficiency of the Nucleotide Excision Repair System Ex Vivo |
title_full_unstemmed | A Method for Assessing the Efficiency of the Nucleotide Excision Repair System Ex Vivo |
title_short | A Method for Assessing the Efficiency of the Nucleotide Excision Repair System Ex Vivo |
title_sort | method for assessing the efficiency of the nucleotide excision repair system ex vivo |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8526188/ https://www.ncbi.nlm.nih.gov/pubmed/34707905 http://dx.doi.org/10.32607/actanaturae.11430 |
work_keys_str_mv | AT popovaa amethodforassessingtheefficiencyofthenucleotideexcisionrepairsystemexvivo AT orishchenkoke amethodforassessingtheefficiencyofthenucleotideexcisionrepairsystemexvivo AT naumenkokn amethodforassessingtheefficiencyofthenucleotideexcisionrepairsystemexvivo AT evdokimovan amethodforassessingtheefficiencyofthenucleotideexcisionrepairsystemexvivo AT petrusevaio amethodforassessingtheefficiencyofthenucleotideexcisionrepairsystemexvivo AT lavrikoi amethodforassessingtheefficiencyofthenucleotideexcisionrepairsystemexvivo AT popovaa methodforassessingtheefficiencyofthenucleotideexcisionrepairsystemexvivo AT orishchenkoke methodforassessingtheefficiencyofthenucleotideexcisionrepairsystemexvivo AT naumenkokn methodforassessingtheefficiencyofthenucleotideexcisionrepairsystemexvivo AT evdokimovan methodforassessingtheefficiencyofthenucleotideexcisionrepairsystemexvivo AT petrusevaio methodforassessingtheefficiencyofthenucleotideexcisionrepairsystemexvivo AT lavrikoi methodforassessingtheefficiencyofthenucleotideexcisionrepairsystemexvivo |