MTK

MTK enables rapid assembly of modular DNA circuits for mammalian cells using Golden Gate-based cloning to construct transcriptional units from a curated parts library for applications including dCas9-based circuit prototyping and generation of noninfectious viral circuits for pharmacological testing.

Key Features:

• Golden Gate-Based Cloning: Uses Golden Gate assembly to combine parts into transcriptional units with high fidelity and scarless junctions.
• Curated Library of Modular Parts: Provides a characterized collection of interchangeable genetic parts for building promoters, coding sequences, and regulatory elements.
• Versatile Assembly Capabilities: Supports construction of single-integration landing pads and generation/delivery of libraries containing protein variants and single-guide RNAs (sgRNAs).
• Iterative Circuit Prototyping: Enables repeated assembly and testing cycles for dCas9-based regulatory circuit development.

Scientific Applications:

• Noninfectious Viral Circuit Generation: Produces noninfectious viral circuits for rapid testing of pharmacological inhibitors against emerging viruses.
• dCas9 Circuit Development: Facilitates prototyping and optimization of dCas9-based gene regulatory circuits in mammalian cells.
• Library-Based Screening: Enables construction and delivery of protein variant and sgRNA libraries for functional screening.
• Cell Engineering with Landing Pads: Supports creation of single-integration landing pads for stable integration of synthetic circuits in mammalian cells.

Methodology:

Selection of modular parts from the curated library followed by assembly into transcriptional units using Golden Gate cloning, with resulting constructs integrated into mammalian cells.