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In Vivo Histone Labeling Using Ultrafast trans-Splicing Inteins
Nicholas A Prescott1,2, Yael David3,4,5,6
1Tri-Institutional PhD Program in Chemical Biology, New York, NY, USA.
Methods in Molecular Biology (Clifton, N.J.)
|March 8, 2020
Summary
Scientists developed a new method for protein trans-splicing (PTS) in live cells. This technique allows for the seamless addition of synthetic or native histone modifications, like fluorophores, directly into chromatin in vivo.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- Expressed protein ligation (EPL) advanced mechanistic biochemistry.
- Protein trans-splicing (PTS) uses split inteins for traceless peptide ligation.
- Existing methods have limitations for in vivo applications.
Purpose of the Study:
- To adapt protein trans-splicing (PTS) methodologies for live-cell applications.
- To enable the delivery of synthetic or native histone modifications in vivo.
- To provide a versatile protocol for chromatin modification.
Main Methods:
- Utilized split inteins for protein trans-splicing.
- Developed a protocol for PTS in live cells.
- Incorporated a small molecule fluorophore into chromatinized histones as an example.
Main Results:
- Successfully adapted PTS for use in live cells.
- Demonstrated the incorporation of a fluorophore into chromatinized histones.
- Established a protocol adaptable for various in vivo chromatin modifications.
Conclusions:
- PTS is a viable method for in vivo chromatin modification.
- The developed protocol facilitates the study of histone modifications in live cells.
- This methodology opens new avenues for mechanistic biochemistry and epigenetics research.
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