High-throughput mapping of modular regulatory domains in human RNA-binding proteins

Area of Science:

• Molecular Biology
• Gene Regulation
• Biochemistry

Background:

• Gene expression is finely tuned by thousands of RNA-binding proteins (RBPs).
• Some RBPs utilize modular domains to recruit regulatory complexes, influencing their function.
• Identifying these RNA-regulatory effector domains is crucial for understanding RBP mechanisms and engineering new RNA regulators.

Purpose of the Study:

• To develop a high-throughput method for identifying RNA-downregulatory effector domains in human RBPs.
• To discover novel RNA-downregulatory effector domains and characterize their function.
• To engineer synthetic RNA regulators for precise control of gene expression.

Main Methods:

• Development of a high-throughput recruitment assay (HT-RNA-Recruit).
• Screening over 30,000 protein tiles from 367 human RBPs against a reporter mRNA.
• Engineering inducible synthetic RNA regulators based on NANOS domains.

Main Results:

• Discovery of over 100 RNA-downregulatory effector domains in 86 human RBPs.
• Identification of domains like KRABs that suppress gene expression when recruited to DNA or RNA.
• Successful engineering of synthetic RNA regulators capable of downregulating endogenous RNAs or maintaining intermediate expression levels.

Conclusions:

• The study provides a valuable resource for understanding RNA regulation and RBP function.
• The identified effector domains expand the toolkit for designing novel RNA-based regulatory systems.
• Engineered synthetic RNA regulators offer precise control over gene expression, with potential applications in biotechnology and medicine.