2026-04-17T03:20:17Zhttps://recercat.cat/oai/request

oai:recercat.cat:2445/2026002026-03-03T03:21:20Zcom_2072_1057col_2072_478902col_2072_478917

00925njm 22002777a 4500 dc Rubio Canalejas, Alba author Pedraz López, Lucas author Torrents Serra, Eduard author 2023-10-06T16:42:53Z 2023-10-06T16:42:53Z 2023-04-18 2023-09-29T09:36:52Z ReViTA (Reverse in VitroTranscription Assay) is a novel in vitro transcription-based method to study gene expression under the regulation of specific transcription factors. The ReViTA system uses a plasmid with a control sequence, the promoter region of the studied gene, the transcription factor of interest, and an RNA polymerase saturated with σ70. The main objective of this study was to evaluate the method; thus, as a proof of concept, two different transcription factors were used, a transcriptional inducer, AlgR, and a repressor, LexA, from Pseudomonas aeruginosa. After the promoters were incubated with the transcription factors, the plasmid was transcribed into RNA and reverse transcribed to cDNA. Gene expression was measured using qRTPCR. Using the ReViTA plasmid, transcription induction of 55% was observed when AlgR protein was added and a 27% transcription reduction with the repressor LexA, compared with the samples without transcription factors. The results demonstrated the correct functioning of ReViTA as a novel method to study transcription factors and gene expression. Thus, ReViTA could be a rapid and accessible in vitro method to evaluate genes and regulators of various species.Crown Copyright © 2023. Published by Elsevier B.V. All rights reserved. Expressió gènica Factors de transcripció Transcripció genètica Gene expression Transcription factors Genetic transcription ReViTA: A novel in vitro transcription system to study gene regulation