PASTMUS

PASTMUS maps functional elements within proteins at single-amino-acid resolution by parsing fragmented DNA sequences from CRISPR tiling mutagenesis to identify residues that influence protein function.

Key Features:

• CRISPR Tiling Mutagenesis Integration: Processes data from CRISPR tiling mutagenesis experiments that generate fragmented DNA sequences covering target proteins.
• Computational Pipeline Integration: Integrates CRISPR-derived sequence fragments with bioinformatics pipelines to associate fragment-level perturbations with functional outcomes.
• Native Biological Context Analysis: Maps functional effects while preserving proteins in their native biological contexts rather than relying on isolated or modified proteins.
• High-Resolution Functional Mapping: Produces amino-acid-resolution functional maps that pinpoint critical residues and their contributions to protein structure and function.

Scientific Applications:

• Protein mapping: High-resolution mapping of six proteins, including three bacterial toxin receptors and three cancer drug targets.
• Mechanistic dissection: Identification of functional residues and residue-level interactions to inform mechanistic studies of protein function.
• Therapeutic target identification: Highlighting residues critical for function to support identification of potential therapeutic targets and guide drug development.

Methodology:

Uses CRISPR tiling mutagenesis to generate libraries of fragmented DNA sequences and parses those fragments with bioinformatics pipelines to identify amino acids that influence protein function.