Monitoring the expression level of coding and non-coding RNAs using a TetR inducing aptamer tag

Meitert, Johannes; Aram, Ronny; Wiesemann, Katharina; Weigand, Julia E.; Suess, Beatrix

doi:10.1016/j.bmc.2013.07.035

Cited by 3 publications

(3 citation statements)

References 29 publications

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“…In some studies, RNA tags did not affect the steady-state levels, regulation or processing of the RNA in which they were embedded (27,28) and in other studies the embedded tags did not affect the cell growth characteristics (28–33). However, some studies have shown that the position of the tag in the RNA, and/or its sequence context, altered the RNA steady-state expression level (34,35).…”

Section: Discussionmentioning

confidence: 99%

Live-cell imaging of Pol II promoter activity to monitor gene expression with RNA IMAGEtag reporters

Shin¹,

Ray²,

Gupta³

et al. 2014

Nucleic Acids Research

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We describe a ribonucleic acid (RNA) reporter system for live-cell imaging of gene expression to detect changes in polymerase II activity on individual promoters in individual cells. The reporters use strings of RNA aptamers that constitute IMAGEtags (Intracellular MultiAptamer GEnetic tags) that can be expressed from a promoter of choice. For imaging, the cells are incubated with their ligands that are separately conjugated with one of the FRET pair, Cy3 and Cy5. The IMAGEtags were expressed in yeast from the GAL1, ADH1 or ACT1 promoters. Transcription from all three promoters was imaged in live cells and transcriptional increases from the GAL1 promoter were observed with time after adding galactose. Expression of the IMAGEtags did not affect cell proliferation or endogenous gene expression. Advantages of this method are that no foreign proteins are produced in the cells that could be toxic or otherwise influence the cellular response as they accumulate, the IMAGEtags are short lived and oxygen is not required to generate their signals. The IMAGEtag RNA reporter system provides a means of tracking changes in transcriptional activity in live cells and in real time.

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Section: Discussionmentioning

confidence: 99%

Live-cell imaging of Pol II promoter activity to monitor gene expression with RNA IMAGEtag reporters

Shin¹,

Ray²,

Gupta³

et al. 2014

Nucleic Acids Research

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“…Another unusual inducer of TetR is the RNA aptamer 12-1. Meitert et al (2013) generated transcriptional fusions resulting in the insertion of the aptamer into untranslated regions of mRNAs as well as into small non-coding RNAs. This served to monitor expression levels of natural transcripts in E. coli.…”

Section: Future Directions Of Tetcontrolfrom Tool To Toolboxmentioning

confidence: 99%

Status quo of tet regulation in bacteria

Bertram

Neumann

Schuster

2021

Microbial Biotechnology

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The tetracycline repressor (TetR) belongs to the most popular, versatile and efficient transcriptional regulators used in bacterial genetics. In the tetracycline (Tc) resistance determinant tet(B) of transposon Tn10, tetR regulates the expression of a divergently oriented tetA gene that encodes a Tc antiporter. These components of Tn10 and of other natural or synthetic origins have been used for tetracycline-dependent gene regulation (tet regulation) in at least 40 bacterial genera. Tet regulation serves several purposes such as conditional complementation, depletion of essential genes, modulation of artificial genetic networks, protein overexpression or the control of gene expression within cell culture or animal infection models. Adaptations of the promoters employed have increased tet regulation efficiency and have made this system accessible to taxonomically distant bacteria. Variations of TetR, different effector molecules and mutated DNA binding sites have enabled new modes of gene expression control. This article provides a current overview of tet regulation in bacteria.

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“…Можно предположить, что принципиальная способность аптамеров как фрагментов ДНК или РНК воспроизводиться в аппарате репликации или транскрипции клетки [76] приведет к разработке аптамеров, включенных в состав плазмидного или фагового генома и способных к высокоэффективному размножению внутри бактерии после ее инфекции фаговой частицей [77] или иным вектором, несущим репликационно-и транскрипционно-компетентный аптамер [78].…”

Section: проблемы селекции и применения аптамеров для создания антибаunclassified

Aptamers in the Treatment of Bacterial Infections: Problems and Prospects

et al. 2016

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Aptamers are short single-stranded oligonucleotides which are selected via targeted chemical evolution in vitro to bind a molecular target of interest. The aptamer selection technology is designated as SELEX (Systematic evolution of ligands by exponential enrichment). SELEX enables isolation of oligonucleotide aptamers binding a wide range of targets of interest with little respect for their nature and molecular weight. A number of applications of aptamer selection were developed ranging from biosensor technologies to antitumor drug discovery. First aptamer-based pharmaceutical (Macugen) was approved by FDA for clinical use in 2004, and since then more than ten aptamer-based drugs undergo various phases of clinical trials. From the medicinal chemist’s point of view, aptamers represent a new class of molecules suitable for the development of new therapeutics. Due to the stability, relative synthesis simplicity, and development of advanced strategies of target specific molecular selection, aptamers attract increased attention of drug discovery community. Difficulties of the development of next-generation antibiotics basing on the conventional basis of combinatorial chemistry and high-throughput screening have also amplified the interest to aptamer-based therapeutic candidates. The present article reviews the investigations focused on the development of antibacterial aptamers and discusses the potential and current limitations of the use of this type of therapeutic molecules.

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Monitoring the expression level of coding and non-coding RNAs using a TetR inducing aptamer tag

Cited by 3 publications

References 29 publications

Live-cell imaging of Pol II promoter activity to monitor gene expression with RNA IMAGEtag reporters

Live-cell imaging of Pol II promoter activity to monitor gene expression with RNA IMAGEtag reporters

Status quo of tet regulation in bacteria

Aptamers in the Treatment of Bacterial Infections: Problems and Prospects

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