Molecular Biology Webinars

Optimizing soluble protein expression: codon optimization, RBS design, and expression vector design

Do you wish there were an easy way to optimize protein expression? This webinar will introduce you to three powerful strategies that can make your research more efficient: codon optimization, ribosome binding site design, and expression vector selection. We will describe the principles underlying these strategies, review case studies showing how big an impact they can have, and walk through how easy it is to use GenScript's free computational tools and best-in-class gene synthesis and cloning services to get optimized expression constructs.

Originally aired November 11, 2015

Codon optimization: Why & how to design DNA sequences for optimal soluble protein expression

Have you struggled with low protein expression levels in your experiments? This webinar will explain the principles of codon optimization and explore case studies showing how it improves protein expression up to 100-fold.  Research has revealed dozens of DNA sequence features that influence the efficiency of each step required to achieve soluble target protein expression. We will review the critical publications that inform GenScript's patented algorithm and the data showing how our algorithm compares to our competitors. We will look at peer-reviewed papers that employed codon-optimized synthetic genes for heterologous protein expression in different host systems, including bacteria, yeast, plant, and human cells. Finally, we will see how GenScript's codon optimization can provide clever solutions to molecular biology problems in specialized applications.

Presented by: Rachel Speer, Ph.D. Originally aired October 29, 2014

Clone less, know more: efficient expression optimization of proteins and pathways using the RBS calculator

Maximizing a heterologous protein's titer can be a balancing act between under- and over-expression. Too much: the protein aggregates. Too little: scale-up is not feasible. This challenge becomes enormous when multiple proteins are involved, for example, when expressing a multi-subunit therapeutic or engineering a multi-enzyme metabolic pathway. It's costly and honestly quite boring to screen combinatorial libraries.

To solve these challenges, this webinar introduces the computational design of synthetic DNA to systematically optimize expression levels in one- or multi-protein genetic systems, while performing the fewest number of experimental measurements. We introduce the physics behind the RBS Calculator, a thermodynamic model that predicts a mRNA's translation initiation rate using only its sequence. We then show how to use the RBS Library Calculator to design small RBS libraries that search large expression spaces. Instead of screening 10^8 variants, our computational design approach requires only 10 to 100 characterized constructs. We present several case studies showing how to optimize protein titers and engineer multi-enzyme metabolic pathways.

Presented by: Prof. Howard Salis, Penn State University

Design high specificity CRISPR/Cas9 gRNAs: principles and tools

The CRISPR/Cas9 system has recently emerged as the most powerful gene editing method to study gene function. This new technology has made gene editing easy to do in any lab. To perform CRISPR/Cas9-mediated gene editing, the first step is to design guide RNA sequences for the target gene. In this webinar, we will present detailed gRNA design principles and provide step-by-step guidance on how to design high specificity gRNAs that avoid off-target effects. We will also introduce the online gRNA design tool and other resources offered by GenScript to help you start doing CRISPR-mediated gene editing with the knowledge and tools you need to be successful.

Presented by: Heidi Huang, Ph.D. Originally aired November 5, 2014

Gene variant libraries: design, construction, and research applications

Do you wish you could enhance your research by using large gene mutant libraries, but find the thought of generating them too daunting? While it can be very challenging and time-consuming to create and validate hundreds or thousands of unique variant sequences in your own lab, GenScript's expertise in gene synthesis technology makes it easy and affordable. In this webinar, we'll discuss different strategies for designing variant libraries, and we'll explore how DNA libraries are being used in peer-reviewed publications from fields including structural biology, vaccine research, synthetic biology, and antibody engineering.

Presented by: Rachel Speer, Ph.D. Originally aired May 21^st and June 18^th, 2014

Building a Synthetic Eukaryotic Genome: Design, Construction and Application

Are you looking for new methods to design and engineer genes, pathways and even chromosomes? Leslie Mitchell, Ph.D., one of the leading scientists working on the Synthetic Yeast genome project Sc2.0 (www.syntheticyeast.org), will present their ambitious work to build the first synthetic eukaryotic genome. Sc2.0 aims to design, construct, and replace the native 12 Mb genome of Saccharomyces cerevisiae with a fully synthetic version. Sc2.0 chromosomes are designed to encode myriad changes. First, to improve genomic stability, destabilizing elements such as transposons are removed and tRNA genes are re-located to a separate 'neochromosome'. Second, synonymously recoded sequences called PCRtags permit encryption and tracking of the synthetic DNA. Finally, to enable downstream genetic flexibility, Sc2.0 encodes an inducible evolution system called SCRaMbLE (Synthetic Chromosome Rearrangement and Modification by LoxP-mediated Evolution) that can generate combinatorial genetic diversity on command. To date, construction has begun on more than half of the 16 synthetic chromosomes by an international team of collaborators. Software and experimental infrastructure developed to facilitate Sc2.0 genome design and construction are applicable to new projects ranging from single gene/pathway design to synthesizing artificial chromosomes.

Presented by: Leslie Mitchell, Ph.D., NYU Langone Medical Center Originally aired June 25^th, 2014

Expression vectors: how to choose, or customize, vectors for gene & protein expression

Do you make new DNA constructs only using the old expression vectors you're most familiar with? This webinar will help you make your experimental design more efficient and powerful by learning how to select or design an expression vector that is optimized for your experiments. We will walk through how to read a plasmid map and what key features to look for in an expression vector depending on your research goal. Using case studies from published literature, we'll discuss why and how you might want to make custom changes to elements already included in commercial available vectors (e.g. RBS or tags). Along with handy reference guides for popular vectors used in different eukaryotic and prokaryotic species, this webinar will introduce you to GenScript's gene synthesis and cloning services that can help you get expression-ready clones most efficiently to accelerate your research.

Presented by: Rachel Speer, Ph.D, Technical Writer, GenScript