A Review of Methods to Monitor the Modulation of mRNA Stability: A Novel Approach to Drug Discovery and Therapeutic Intervention

Abstract

Posttranscriptional regulation of gene expression is an elaborate and intricate process, constituting an important mechanism for the control of protein expression. During its existence, mRNA is escorted by proteins and other RNAs, which control the maturation, transportation, localization, translational efficiency, and ultimately its degradation. Without changes at the transcription level, mRNA steady-state levels can vary dramatically by just small changes in mRNA stability. By influencing the metabolism of specific mRNAs, the abundance of specific mRNAs can be controlled in organisms from bacteria to mammals. In eukaryotic cells, the control of mRNA stability is exerted through specific cis-acting elements (sequence-specific control elements) and trans-acting factors (mRNA binding proteins and some miRNAs). mRNA stability appears to be a key regulator in controlling the expression of many proteins. Dysregulation of mRNA stability has been associated with human diseases, including cancer, inflammatory disease, and Alzheimer's. These observations suggest that modulating the stability of specific mRNAs may represent a viable strategy for pharmaceutical intervention. The literature already describes several compounds that influence mRNA stability. Measuring mRNA stability by conventional methods is labor intensive and time-consuming. However, several systems have been described that can be used to screen for modulators of mRNA levels in a high-throughput format. Thus, these assay systems offer a novel approach for screening targets that at present appear to be poorly "drugable." This review describes the utility of mRNA stability as a novel approach to drug discovery, focusing on assay methods and tool compounds available to monitor mRNA stability. The authors describe mRNA stability assays and issues related to this approach.

Tristetraprolin-dependent Post-transcriptional Regulation of Inflammatory Cytokine mRNA Expression by Apolipoprotein A-I

“…Our studies provide evidence that the suppressive function of apoA-I in inflammation is mediated, at least partially, by antagonism of LPS-induced inflammatory genes in mRNA stability. It has been reported that dysregulation of mRNA stability is associated with some pathologies such as chronic inflammation and cardiovascular diseases (39,40). LPS, a classic stimulator of Toll-like receptor 4 (TLR4)-mediated inflammation, has also been reported to promote stabilization of pro-inflammatory mRNAs such as TNF-␣ and IL-6 (41, 42).…”

Sp6 regulation of <i>Rock1 </i>promoter activity in dental epithelial cells

“…It remains unclear whether other regions in Rock1 promoter are involved in Sp1-mediated endogenous promoter activity. Furthermore, we can measure the steady-state levels of mRNA by RT-PCR analysis, which are regulated at the transcriptional level and stability in a post-transcriptional manner (19). The mRNA stability is regulated by the interaction between cis-acting, i.e.…”

Influência do hormônio tireoideano na atividade do gonadotrofo.

“…A extração do RNA total foi baseada na metodologia da guanidina-fenolclorofórmio e precipitação com isopropanol (CHOMCZYNSKI et al, 1987 …”

“…As amostras de testículos e epidídimos foram pulverizadas em nitrogênio líquido e uma porção (30 mg de tecido) depositada num tubo tipo eppendorff de 2,0 mL e submetida à extração do RNA total, baseada na metodologia da guanidina-fenol-clorofórmio e precipitação com isopropanol (CHOMCZYNSKI et al, 1987), conforme procedimentos já descritos previamente.…”

“…Deste modo, a regulação do turnover do mRNA é uma etapa essencial para a avaliação da sua abundância e, portanto, da expressão gênica (CHENEVAL et al, 2010 A actina realiza um processo dinâmico no qual a forma monomérica (actina globular ou Gactina) se polimeriza em filamentos (actina filamentosa ou F-actina), sendo este processo reversível (MIRALES e VISA, 2006;FIRAT-KARALAR e WELCH, 2011). Dessa forma, a actina participa de processos envolvidos com a transcrição gênica, já que direciona fatores transcricionais ao núcleo, bem como se relaciona a eventos traducionais, ancorando mRNAs e fatores de alongamento, direcionando-os à maquinaria de tradução (GOULART-SILVA et al, 2006;SILVA et al, 2010).…”