Characterizing protein sequence determinants of nuclear condensates by high-throughput pooled imaging with CondenSeq

Area of Science:

• Cell Biology
• Biochemistry
• Genomics

Background:

• Biomolecular condensates are essential for cellular organization and function.
• Dysregulation of condensates is linked to neurodegenerative diseases and cancer.
• Understanding sequence determinants of condensate formation is critical.

Purpose of the Study:

• To systematically explore sequence features governing protein condensate formation.
• To develop a high-throughput method for measuring condensate propensity.
• To identify sequence properties driving different condensate classes.

Main Methods:

• Developed CondenSeq: a pooled imaging with in situ sequencing approach.
• Measured condensate formation propensity for thousands of protein sequences.
• Analyzed sequence features across diverse contexts.

Main Results:

• Identified sequence features with consistent and context-dependent effects on condensate formation.
• Discovered distinct sequence properties for multiple condensate classes.
• Established a quantitative relationship between sequence and condensate propensity.

Conclusions:

• Protein sequence intrinsically dictates nuclear condensate formation.
• CondenSeq provides a scalable platform for dissecting sequence-structure-function relationships.
• Findings advance understanding of condensate biology and disease implications.