MuSi

MuSi enables analysis and design of codon-directed mutagenesis gene libraries to control amino acid types and distributions using trinucleotide mixtures and to evaluate library diversity.

Key Features:

• Codon-directed mutagenesis analysis: Analyzes gene libraries that have undergone codon-directed mutagenesis to support variant discovery.
• Trinucleotide mixture control: Supports the use of trinucleotide mixtures for controlled randomization to specify amino acid types and distributions at each position.
• Combinatorial assembly strategy: Implements a combinatorial assembly approach to create full-length gene libraries from smaller sub-libraries.
• Sequence-based diversity evaluation: Provides comprehensive sequence analysis capabilities for efficient assessment of library diversity.
• Targeted codon randomization: Enables targeting of selected codons with integrated rational criteria and contingency to enhance diversity.
• Library scalability and preservation: Facilitates on-demand assembly of high-diversity libraries from less diverse components to address archiving and repeated propagation issues with phages or plasmids.

Scientific Applications:

• High-diversity library generation: Generates high-diversity gene libraries using trinucleotide-directed randomization.
• Protein variant discovery: Enables exploration and selection of functional protein variants from combinatorial libraries.
• Scaffold engineering (tHisF): Applied to the tHisF scaffold from Thermotoga maritima to target 26 selected codons for controlled randomization.
• Stability and activity assessment: Supports assessment of conformational stability of library members and isolation of variants exhibiting HisF activity.
• Mitigation of propagation issues: Supports strategies to reduce problems associated with long-term archiving and repeated proliferation of phages or plasmids.

Methodology:

Provides sequence analysis of mutated libraries to evaluate library composition and diversity.