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Purification of Single-Stranded RNA Bacteriophages and Host Receptors for Structural Determination Using Cryo-Electron Microscopy
Single-stranded RNA bacteriophages (ssRNA phages) are small viruses with a compact genome (~3–4 kb) that infect gram-negative bacteria via retractile... -
Protocols for Building and Producing High Diversity Peptide Phage Display Libraries
Phage display is an important technology to study protein-protein interaction and protein evolution, with applications in basic science and applied... -
Generation of a Naïve Human scFv Phage Display Library and Panning Selection
Phage display antibody libraries have been successfully used as the essential tool to produce monoclonal antibodiesMonoclonal antibodies (mAB)... -
Combining a Base Deaminase Mutator with Phage-Assisted Evolution
Phage-assisted evolution has emerged as a powerful technique for improving a protein’s function by using mutagenesis and selective pressure. However,... -
Plant Expression of Trans-Encapsidated Chimeric Viral Vaccines with Animal RNA Replicons: An Update
In this protocol, we outline how to produce a chimeric viral vaccine in a biosafety level 1 (BSL1) environment. An animal viral vector RNA... -
Trans-Amplifying RNA Vaccines Against Infectious Diseases: A Comparison with Non-Replicating and Self-Amplifying RNA
The recent COVID-19 pandemic as well as other past and recent outbreaks of newly or re-emerging viruses show the urgent need to develop potent new... -
Sequence-Optimized mRNA Vaccines Against Infectious Disease
Develo** effective mRNA vaccines poses certain challenges concerning mRNA stability and ability to induce sufficient immune stimulation and... -
Multiplex Functional Characterization of Protein Variant Libraries in Mammalian Cells with Single-Copy Genomic Integration and High-Throughput DNA Sequencing
Sequencing-based, massively parallel genetic assays have enabled simultaneous characterization of the genotype–phenotype relationships for libraries... -
High-Throughput Spectroscopic Analysis of mRNA Cap** Level
Eukaryotic mRNAs are characterized by terminal 5′ cap structures and 3′ polyadenylation sites, which are essential for posttranscriptional... -
Detection of MicroRNAs Using Synthetic Toehold Switch in Mammalian Cells
Engineering synthetic gene circuits to control cellular functions has a broad application in the field of synthetic biology. Synthetic RNA-based... -
A Computational Modeling Approach for the Design of Genetic Control Systems that Respond to Transcriptional Activity
Recent progress in synthetic biology has enabled the design of complex genetic circuits that interface with innate cellular functions, such as gene... -
A Directed Evolution Protocol for Engineering Minimal Transcription Factors, Based on CIS Display
Directed evolution is an efficient strategy for obtaining desired biomolecules. Since the 1990s, the emergence of display techniques has enabled... -
Imaging S-Adenosyl Methionine Dynamics in Living Cells Using an RNA-Based Fluorescent Sensor
S-Adenosyl methionine (SAM) is a critical metabolite involved in numerous cellular processes, including DNA methylation and gene expression... -
Posttranslational Remote Control Mediated by Physical Inducers for Rapid Protein Release in Engineered Mammalian Cells
Physical cues such as light, heat, or an electrical field can be utilized for traceless, on-demand activation of the expression of a desired... -
Coiled-Coil Interaction Toolbox for Engineering Mammalian Cells
Protein interactions play a crucial role in a variety of biological processes. Therefore, regulation of these interactions has received considerable... -
Genome Editing, Transcriptional Regulation, and Forward Genetic Screening Using CRISPR-Cas12a Systems in Yarrowia lipolytica
Class II Type V endonucleases have increasingly been adapted to develop sophisticated and easily accessible synthetic biology tools for genome... -
Multiplex Marker-Less Genome Integration in Pichia pastoris Using CRISPR/Cas9
Pichia pastoris is known for its excellent protein expression ability. As an industrial methyl nutritional yeast, it can effectively utilize methanol... -
Cell-Free Synthesis and Quantitation of Bacteriophages
Cell-free transcription-translation (TXTL) enables achieving an ever-growing number of applications, ranging from the rapid characterization of DNA... -
Genetic Network Design Automation with LOICA
Genetic design automation (GDA) is the use of computer-aided design (CAD) in designing genetic networks. GDA tools are necessary to create more... -
The Causes for Genomic Instability and How to Try and Reduce Them Through Rational Design of Synthetic DNA
Genetic engineering has revolutionized our ability to manipulate DNA and engineer organisms for various applications. However, this approach can lead...