Titlebook: Riboregulator Design and Analysis; James Chappell,Melissa K. Takahashi Book 2022 The Editor(s) (if applicable) and The Author(s), under ex

Ovulation

Development of Synthetic Riboswitches to Guide the Evolution of Metabolite Production in Microorganregulate the expression of a gene or an operon depending on the intracellular level of a target molecule and consequently modulate the cellular responses. In evolutionary engineering, riboswitch-based biosensors have been widely applied for high-throughput screening or selection of target phenotypes

Acetabulum

Efficient Method to Identify Synthetic Riboswitches Using RNA-Based Capture-SELEX Combined with In based switches have to exhibit excellent binding affinity as well as ligand binding-induced structural changes. Selection via Capture-SELEX favors the enrichment of aptamers which exhibit both characteristics. For the Capture-SELEX, an RNA pool is used that gets immobilized onto a capture oligonucle

破裂

RNA Design Principles for Riboswitches that Regulate RNase P-Mediated tRNA Processing, an aptamer platform for the highly selective ligand recognition and an expression platform which controls gene activity typically at the level of transcription or translation. In previous work, we could successfully apply RNA folding prediction to implement a new riboswitch mechanism regulating pro

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Design, Characterization, and Application of Targeted Gene Activation in Bacteria Using a Modular Cof the single guide RNA (sgRNA) to target DNA sequences. Included in this are CRISPR activation (CRISPRa) systems. These systems allow users to activate transcription of a target gene through the localization of transcription activation domains (ADs) near promoter elements, which in turn recruit RNA

微生物

Reprogramming TracrRNAs for Multiplexed RNA Detection,e how our recent discovery of noncanonical crRNAs inspired the engineering of reprogrammed tracrRNAs and led to a powerful platform for multiplexed RNA detection. We provide detailed protocols regarding how to design reprogrammed tracrRNA and carry out?assays in vitro and in vivo.

嬰兒

Harnessing CRISPR-Cas9 for Epigenetic Engineering, promoters and enhancers. Here, we describe a basic methodology for performing an epigenome editing experiment, starting from gRNA design and cloning to transiently transfecting the gRNA plasmid and the CRISPR/dCas9-based epigenetic effector and finalizing with chromatin immunoprecipitation (ChIP) t

機制

RNA Structure Prediction, Analysis, and Design: An Introduction to Web-Based Tools,utational algorithms that utilize thermodynamics have been developed to predict RNA secondary structure. Due to the importance of intermolecular interactions, the algorithms have been expanded to determine and predict RNA-RNA hybridization. This chapter discusses popular webservers with the tools fo

匍匐前進

Single-Molecule FRET Studies of RNA Structural Rearrangements and RNA-RNA Interactions, its target are typically facilitated by a protein partner. Single-molecule fluorescence microscopy is a powerful tool for dissecting the conformational states and intermediates that contribute to target recognition. This chapter describes protocols for studying target recognition by bacterial small

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Laconic

Computational Design of Small Transcription Activating RNAs (STARs),of terminator hairpins to activate the transcription of targeted genes. STARs have found broad utility for creating synthetic gene circuits, engineering metabolic pathways, and creating new types of diagnostics. Here we describe the method to computationally design, build, and characterize STAR regulators.