UCLA researchers from the Department of Bioengineering have developed artificial RNA sequences that can form condensates in vitro, allowing customization of cellular function in membrane-less organelles.
BACKGROUND: RNA condensates are intracellular membrane-less compartments formed by phase separation, involving RNA and proteins. These RNA condensates play integral roles in various cellular functions, including transcriptional regulation and protection from RNA degradation. Additionally, these condensates are thought to play a role in diseases such as neurodegeneration. Because of the importance of RNA condensates to cellular function, the ability to synthetically program and customize RNA condensation is extremely valuable in allowing researchers to program cellular behavior.
INNOVATION: UCLA researchers have developed artificial, modular RNA sequences that form phase separated condensates, termed RNA nanostars. As these RNA sequences are transcribed, they form modular stem-loop domains that assemble via programmable base-pairing interactions. Since these sequences can be optimized and diversified, it is possible to generate distinct RNA condensates that recruit other small molecules, RNA, and proteins to these organelles to catalyze biochemical reactions and even potentially improve cell fitness. Additionally, the researchers developed methods to dissolve theses RNA condensates by inducing a conformational change that prevents condensation, allowing researchers to powerfully manipulate these condensates. Furthermore, this novel technology allows RNA condensates to be synthesized directly from a DNA template, allowing them to be able to be formed in living cells. RNA nanostars have the potential to manipulate cellular behavior, compartmentalize small molecules for biochemical reactions, and develop treatments for diseases.
POTENTIAL APPLICATIONS:
- Compartmentalization of cellular reactions in synthetic and biological cells
- Potential as platform technology for diagnostic applications, smart drug delivery system, and purification system
ADVANTAGES:
- Generation of RNA condensates from a DNA template allows endogenous cellular machinery to produce condensates in living cells.
- RNA sequences can be customized, allowing multiple orthogonal compartments to be formed within one cell.
DEVELOPMENT-TO-DATE: UCLA researchers have developed customizable RNA sequences that assemble into RNA condensates in vitro, as well as developed methods to dissolve these condensates. They are currently adapting these RNA condensates to be produced in living cells.
Follow-up patent application: E. Franco, S. Li, and A. Tang. “Methods for building artificial RNA organelles in living cells”. U.S. Provisional Patent Application No. 63/679,560. 2024.
Related Papers (from the inventors only):
- Stewart, J. M. et al. Modular RNA motifs for orthogonal phase separated compartments. Nature Communications 15, (2024)
- Fabrini, G. et al. Co-transcriptional production of programmable RNA condensates and synthetic organelles. Nature Nanotechnology (2024). doi:10.1038/s41565-024-01726-x
Keywords: RNA condensates, phase separation, membrane-less organelles, synthetic biology, nanotechnology, constructs, orthogonal compartmentalization, RNA nanostructures