TY - JOUR

T1 - A Method for Assessing the Efficiency of the Nucleotide Excision Repair System Ex Vivo

AU - Popov, A. A.

AU - Orishchenko, K. E.

AU - Naumenko, K. N.

AU - Evdokimov, A. N.

AU - Petruseva, I. O.

AU - Lavrik, O. I.

N1 - Funding Information: This study was supported by the Russian Science Foundation (project No. 19-74-10056); acquisition and analysis of images were partially supported by the budget project No. 0259-2021-0011. Publisher Copyright: © 2021 National Research University Higher School of Economics. All Rights Reserved.

PY - 2021/9

Y1 - 2021/9

N2 - 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.

AB - 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.

KW - DNA damages

KW - ex vivo methods

KW - nucleotide excision repair

UR - http://www.scopus.com/inward/record.url?scp=85121271381&partnerID=8YFLogxK

UR - https://elibrary.ru/item.asp?id=47379995

UR - https://www.mendeley.com/catalogue/f4bfd47f-8118-3d5a-b486-aa62fe29a595/

U2 - 10.32607/actanaturae.11430

DO - 10.32607/actanaturae.11430

M3 - Article

C2 - 34707905

AN - SCOPUS:85121271381

VL - 13

SP - 122

EP - 125

JO - Acta Naturae

JF - Acta Naturae

SN - 2075-8251

IS - 3

M1 - 13

ER -