Genome Editing
Genome Editing
Application
Benchling for CRISPR Screening
CRISPR screening is a revolutionary methodology to identify genes of interest underlying a specific phenotype. CRISPR can be scaled up for screening applications because of the versatility of CRISPR/Cas9 in disrupting target DNA and the ease of gRNA/plasmid generation. However, large-scale use of th... More
Protocol
Lentiviral titering by crystal violet staining
This protocol provides an example for titering lentiviral particles using crystal violet staining in a 6-well format to obtain transducing units per mL (TU/mL). Once lentiviral particles are packaged and collected, small aliquots should be frozen at –80°C. One aliquot should be thawed on ice and use... More
Protocol
Multiplexed, trackable CRISPR-based genome engineering for optimization of heterologous protein expression and activity in E. coli
This work demonstrates the use of the Onyx platform for interrogation and optimization of heterologous proteins in E. coli. Compared to traditional methods for protein engineering, such as random mutagenesis or targeted site-directed mutagenesis approaches, the Onyx technology dramatically increases... More
Protocol
Massively parallel genome engineering followed by pooled growth selections for rapid target discovery in microbes
The ability to engineer the entire genome of microorganisms at a massively parallel scale using the Onyx platform will reshape the manner in which researchers perform pooled cultivation experiments (such as ALE) and generate data sets. Here, Onyx technology was used to rapidly generate genomewide k... More
Protocol
Dharmacon Edit-R crRNA Libraries
Libraries of predesigned synthetic crRNAs provide an opportunity to apply the CRISPR-Cas9 system for functional gene knockout analysis, or dCas9-VPR system for transcriptional gene activation, in an arrayed format. Dharmacon crRNA libraries for functional gene knockout consist of Edit-R synthetic cr... More
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