@inbook{ea1590840b6a4a5eb7fdde7028137634,
title = "CRISPR/Cas9 to Silence Long Non-Coding RNAs",
abstract = "Knockout (KO) of long non-coding RNAs (lncRNAs) enables functional characterization of this still poorly described group of transcripts. One of the most efficient and simplest methods to achieve complete KO of a lncRNA is by employing CRISPR/Cas gene editing. As most lncRNAs are not well annotated, their individual functional regions are often not defined, and the majority of the transcripts are not affected by single nucleotide mutations. Therefore, CRISPR/Cas KO is more challenging for lncRNAs as compared to KO of protein coding genes. Strategies for lncRNAs KO include complete removal of the entire gene, removal of the promoter and transcriptional start site, abolishing exon–exon junctions, or removing the transcriptional termination site. Here, we describe the methodology to perform CRISPR/Cas9 KO of lncRNAs in vitro using electroporation as the method of transfection of presynthesized single guide RNAs (sgRNAs) and Cas9 enzyme.",
keywords = "CRISPR/Cas9, Electroporation, Gene editing, Knockout, Long non-coding RNA",
author = "Rosenlund, {Ingrid Arctander} and Calin, {George A.} and Dragomir, {Mihnea P.} and Erik Knutsen",
note = "Publisher Copyright: {\textcopyright} 2021, Springer Science+Business Media, LLC, part of Springer Nature.",
year = "2021",
doi = "10.1007/978-1-0716-1581-2_12",
language = "English (US)",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "175--187",
booktitle = "Methods in Molecular Biology",
}