Affinity-Based Interactome Analysis of Endogenous LINE-1 Macromolecules.

Autor:	Di Stefano LH; European Research Institute for the Biology of Ageing, University Medical Center Groningen, Groningen, Netherlands., Saba LJ; European Research Institute for the Biology of Ageing, University Medical Center Groningen, Groningen, Netherlands., Oghbaie M; Laboratory of Cellular and Structural Biology, The Rockefeller University, New York, NY, USA., Jiang H; Laboratory of Cellular and Structural Biology, The Rockefeller University, New York, NY, USA., McKerrow W; Institute for Systems Genetics, NYU Langone Health, New York, NY, USA., Benitez-Guijarro M; GENYO. Centro de Genómica e Investigación Oncológica: Pfizer-Universidad de Granada-Junta de Andalucía, Granada, Spain., Taylor MS; Department of Pathology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA., LaCava J; European Research Institute for the Biology of Ageing, University Medical Center Groningen, Groningen, Netherlands. j.p.lacava@rug.nl.; Laboratory of Cellular and Structural Biology, The Rockefeller University, New York, NY, USA. j.p.lacava@rug.nl.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2023; Vol. 2607, pp. 215-256.
DOI:	10.1007/978-1-0716-2883-6_12
Abstrakt:	During their proliferation and the host's concomitant attempts to suppress it, LINE-1 (L1) retrotransposons give rise to a collection of heterogeneous ribonucleoproteins (RNPs); their protein and RNA compositions remain poorly defined. The constituents of L1-associated macromolecules can differ depending on numerous factors, including, for example, position within the L1 life cycle, whether the macromolecule is productive or under suppression, and the cell type within which the proliferation is occurring. This chapter describes techniques that aid the capture and characterization of protein and RNA components of L1 macromolecules from tissues that natively express them. The protocols described have been applied to embryonal carcinoma cell lines that are popular model systems for L1 molecular biology (e.g., N2102Ep, NTERA-2, and PA-1 cells), as well as colorectal cancer tissues. N2102Ep cells are given as the use case for this chapter; the protocols should be applicable to essentially any tissue exhibiting endogenous L1 expression with minor modifications. (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze:	MEDLINE
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