A longstanding goal of synthetic biology has been the programmable control of cellular functions. Central to this is the creation of versatile regulatory toolsets that allow for programmable control of gene expression. Of the many regulatory molecules available, RNA regulators offer the intriguing possibility of de novo design-allowing for the bottom-up molecular-level design of genetic control systems. Here we present a computational design approach for the creation of a bacterial regulator called Small Transcription Activating RNAs (STARs) and create a library of high-performing and orthogonal STARs that achieve up to ~ 9000-fold gene activation. We demonstrate the versatility of these STARs-from acting synergistically with existing constitutive and inducible regulators, to reprogramming cellular phenotypes and controlling multigene metabolic pathway expression. Finally, we combine these new STARs with themselves and CRISPRi transcriptional repressors to deliver new types of RNA-based genetic circuitry that allow for sophisticated and temporal control of gene expression.
RBSDesigner: software for designing synthetic ribosome binding sites that yields a desired level of protein expression
De novo automated design of small RNA circuits for engineering synthetic riboregulation in living cells
Predictive design of mRNA translation initiation region to control prokaryotic translation efficiency
Characterization of 582 natural and synthetic terminators and quantification of their design constraints
Protocols for implementing an Escherichia coli based TX-TL cell-free expression system for synthetic biology
Advances in genetic circuit design: novel biochemistries, deep part mining, and precision gene expression
Rapidly characterizing the fast dynamics of RNA genetic circuitry with cell-free transcription-translation (TX-TL) systems
Engineering modular and tunable genetic amplifiers for scaling transcriptional signals in cascaded gene networks
A predictive biophysical model of translational coupling to coordinate and control protein expression in bacterial operons
Improving fold activation of small transcription activating RNAs (STARs) with rational RNA engineering strategies
Design and Construction of Generalizable RNA-Protein Hybrid Controllers by Level-Matched Genetic Signal Amplification
Dynamic regulation of metabolic flux in engineered bacteria using a pathway-independent quorum-sensing circuit
Determining the Transcription Rates Yielding Steady-State Production of mRNA in the Lac Genetic Switch of Escherichia coli
Next-level riboswitch development-implementation of Capture-SELEX facilitates identification of a new synthetic riboswitch
Development of a novel strategy for robust synthetic bacterial promoters based on a stepwise evolution targeting the spacer region of the core promoter in Bacillus subtilis
Analyzing genomic data using tensor-based orthogonal polynomials with application to synthetic RNAs.
Rational engineering of a modular bacterial CRISPR-Cas activation platform with expanded target range.
Recent advances in tuning the expression and regulation of genes for constructing microbial cell factories.
Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). CRISPR-Cas system enables the editing of genes to create or correct mutations. Discover the latest research on CRISPR here.
CRISPR for Genome Editing
Genome editing technologies enable the editing of genes to create or correct mutations. Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). Here is the latest research on the use of CRISPR-Cas system in gene editing.