@inbook{a5cf34906f584c5ea3c9a8c3ce0ec526,
title = "Characterization of L1-ribonucleoprotein particles",
abstract = "The LINE-1 retrotransposon (L1) encodes two proteins, ORF1p and ORF2p, which bind to the L1 RNA in cis, forming a ribonucleoprotein (RNP) complex that is critical for retrotransposition. Interactions with both permissive and repressive host factors pervade every step of the L1 life cycle. Until recently, limitations in detection and production precluded in-depth characterization of L1 RNPs. Inducible expression and recombinant engineering of epitope tags have made detection of both L1 ORFs routine. Here, we describe large-scale production of L1-expressing HEK-293T cells in suspension cell culture, cryomilling and affinity capture of L1 RNP complexes, sample preparation for analysis by mass spectrometry, and assay using the L1 element amplification protocol (LEAP) and qRT-PCR.",
keywords = "Affinity purification, Cryomilling, Interactomics, LINE-1, Mass spectrometry, Metabolic labelling, Protein complexes, Ribonucleoprotein",
author = "Taylor, {Martin S.} and {La Cava}, John and Lixin Dai and Paolo Mita and Burns, {Kathleen H.} and Rout, {Michael P.} and Boeke, {Jef D.}",
note = "Publisher Copyright: {\textcopyright} Springer Science+Business Media New York 2016.",
year = "2016",
doi = "10.1007/978-1-4939-3372-3_20",
language = "English (US)",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "311--338",
booktitle = "Methods in Molecular Biology",
}