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Protocol for in vitro phase separation of ARID3B proteins
Wenyu Han1, Xiaofeng Li1, Junyan Lin1
1Department of Orthodontics, The Affiliated Stomatological Hospital of Nanjing Medical University, Nanjing, China; State Key Laboratory Cultivation Base of Research, Prevention and Treatment for Oral Diseases (Nanjing Medical University), Nanjing, China; Jiangsu Province Engineering Research Center of Stomatological Translational Medicine (Nanjing Medical University), Nanjing, China.
Researchers developed a protocol to visualize liquid-liquid phase separation (LLPS) of ARID3B, a key factor in cleft lip and palate. This method allows studying ARID3B
Area of Science:
- Biochemistry
- Molecular Biology
- Developmental Biology
Background:
- Liquid-liquid phase separation (LLPS) is increasingly recognized as a crucial mechanism in cellular organization and function.
- Aberrant LLPS of specific proteins, such as ARID3B, has been implicated in developmental disorders like nonsyndromic cleft lip/palate.
Purpose of the Study:
- To present a detailed protocol for visualizing ARID3B liquid-liquid phase separation (LLPS).
- To enable the study of ARID3B phase separation dynamics in vitro and in vivo.
- To provide a framework for investigating LLPS in other transcription factors.
Main Methods:
- Purification of ARID3B protein.
- In vitro regeneration of ARID3B liquid-liquid phase separation (LLPS).
- Detection of ARID3B condensates within mammalian cells.
Main Results:
- A reproducible protocol for visualizing ARID3B LLPS was established.
- The protocol allows for the observation of ARID3B condensate formation and dynamics.
- Successful detection of ARID3B condensates in cellular contexts.
Conclusions:
- The developed protocol provides a valuable tool for analyzing ARID3B phase separation.
- This method facilitates research into the role of ARID3B LLPS in nonsyndromic cleft lip/palate.
- The protocol can be adapted to study phase separation of other transcription factors.
