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System and method for light-regulated oligomerization and phase separation of folded domains and RNA granule-associated protein domains

A protein and domain technology, applied in chemical instruments and methods, DNA/RNA fragments, recombinant DNA technology, etc., which can solve problems such as poorly understood biophysical functions of modular architectures

Pending Publication Date: 2021-11-09
THE TRUSTEES FOR PRINCETON UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the mechanisms by which G3BP regulates SG biogenesis and the biophysical roles of its modular architecture remain poorly understood (Kedersha et al., 2016; Matsuki et al., 2013; Panas et al., 2015; Schulte et al., 2016; Solomon et al., 2007; Wu et al. , 2016)

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  • System and method for light-regulated oligomerization and phase separation of folded domains and RNA granule-associated protein domains
  • System and method for light-regulated oligomerization and phase separation of folded domains and RNA granule-associated protein domains
  • System and method for light-regulated oligomerization and phase separation of folded domains and RNA granule-associated protein domains

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Embodiment Construction

[0056] specific implementation plan

[0057] The present disclosure relates to systems and methods for light-regulated oligomerization and phase separation of fold domains and RNA particle-associated protein domains, particularly for drug-based screening applications.

[0058] The system can involve multiple types of fusion proteins, any or all of which contain fluorescent proteins. These fusion proteins are configured to work together to co-oligomerize and network when illuminated with certain wavelengths of light. This oligomerization and networking (or lack thereof) results in specific phase behavior which can be monitored under various conditions and circumstances (e.g. when various chemical or biological reagents are added) to determine under what conditions or circumstances the phase Behavior can be changed.

[0059] The simplest form of the system can be found in Figures 1A-1C . refer to Figure 1A and 1B , the system requires one or more first fusion proteins (1...

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Abstract

Disclosed is a method and system for the phase separation of folded domains, and more particularly, to inducing clusters of folded domains as part of a drug-based screening application. The system and method utilize one or more first fusion proteins (100, 101), each first fusion protein comprising a first region (110) fused to a second region (120), the first region (110) comprising at least one light sensitive protein (115) or cognate partner of a light sensitive protein (116), and the second region (120) comprising one or more folded RNA binding domains (RBDs), disordered RBDs, folded non-RBD domains, or combination thereof (125).

Description

[0001] Cross References to Related Applications [0002] This application claims priority to U.S. Provisional Application No. 62 / 807459, filed February 19, 2019, the entire contents of which are incorporated herein by reference. technical field [0003] The present disclosure relates generally to phase separation of folded domains, and more particularly to the induction of clusters of folded domains as part of drug-based screening applications. Background technique [0004] Cells compartmentalize the diverse biochemical processes necessary for life in reaction centers called organelles. Organelles are often described as membrane-enclosed compartments isolated from a homogeneous cytosolic solution. However, cells also use membrane-lacking organelles to organize their contents. Such compartments are particularly abundant in the nucleus of eukaryotic cells and include ribosome-producing nucleoli as well as RNA-protein bodies of unknown function (eg Cajal bodies, speckles) (Z...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12N15/115C12N15/63
CPCC07K14/47C07K14/415C07K2319/85C07K2319/735C12N9/22C12N15/113C12N15/62C12N15/635G01N33/5035G01N33/542
Inventor C·P·布朗温内D·布拉查V·德拉克D·W·桑德斯
Owner THE TRUSTEES FOR PRINCETON UNIV