In vivo histone labeling using ultrafast trans-splicing inteins Journal Article
| Authors: | Prescott, N. A.; David, Y. |
| Article Title: | In vivo histone labeling using ultrafast trans-splicing inteins |
| Abstract: | The development of expressed protein ligation (EPL) widened the scope of questions that could be addressed by mechanistic biochemistry. Protein trans-splicing (PTS) relies on the same basic chemical principles, but utilizes split inteins to tracelessly ligate distinct peptide or polypeptide fragments together with native peptide bonds. Here we present a method to adapt PTS methodologies for their use in live cells, in order to deliver synthetic or native histone modifications. As an example, we provide a protocol to incorporate a small molecule fluorophore into chromatinized histones. The protocol should be easily adaptable to incorporate other modifications to chromatin in vivo. © 2020, Springer Science+Business Media, LLC, part of Springer Nature. |
| Keywords: | protein expression; in vivo study; molecular cloning; amino acid sequence; genetic transfection; histone; chromatin; western blotting; protein transport; cell fractionation; amino acid; disulfide; peptide synthesis; conjugation; solid; synthetic peptide; histone modification; synthetic biology; trans splicing; chemical labeling; intein; live cell imaging; in cellulo labeling; split inteins |
| Journal Title: | Methods in Molecular Biology |
| Volume: | 2133 |
| ISSN: | 1064-3745 |
| Publisher: | Humana Press Inc |
| Date Published: | 2020-01-01 |
| Start Page: | 201 |
| End Page: | 219 |
| Language: | English |
| DOI: | 10.1007/978-1-0716-0434-2_10 |
| PUBMED: | 32144669 |
| PROVIDER: | scopus |
| PMCID: | PMC7534146 |
| DOI/URL: | |
| Notes: | Book Chapter 10 in "Expressed Protein Ligation: Methods and Protocols" Vila-Perello M, ed (ISBN: 978-1-0716-0433-5) -- Export Date: 1 April 2020 -- Source: Scopus |
