FRAGLER

FRAGLER automates modular DNA assembly and fragment design to streamline plasmid construct generation for applications such as reporter expression (sfGFP, sfCherry3C) in E. coli, SARS-CoV-2 protein reagent production, and construction of chimeric antigen receptor (CAR) variant libraries.

Key Features:

• Modular Cloning Platform: Multimodule Golden Gate–based cloning framework supports generation of complex constructs and validated splitting and expression of cassettes such as sfGFP and sfCherry3C in E. coli with fluorometric assessment of assembly efficiency.
• Automated Fragment Generation: Automates codon optimization, multiple sequence alignment, and automated sequence fragmentation to generate recyclable fragments for assembly, reducing timelines and costs.
• Flexibility and Robustness: Enables generation of plasmids for SARS-CoV-2 protein reagents, automated parallel assemblies, and creation of modular libraries of chimeric antigen receptor (CAR) variants.
• Scalable Assembly Framework: Provides a scalable assembly framework to streamline high-throughput plasmid construction workflows.

Scientific Applications:

• High-throughput library construction: Generation of modular and combinatorial CAR variant libraries and other plasmid libraries for screening.
• Recombinant protein reagent production: Assembly of plasmids for SARS-CoV-2 protein reagents.
• Reporter expression and assay validation: Construction and expression of reporter cassettes (sfGFP, sfCherry3C) in E. coli with fluorometric assessment.
• Drug discovery and synthetic biology workflows: Scalable plasmid assembly to support drug discovery and synthetic biology applications.

Methodology:

Performs codon optimization, multiple sequence alignment, automated sequence fragmentation and fragment recycling, and orchestration of automated parallel assemblies.