@inbook{4cc27d727d854624a20aa8def85df6a6,
title = "CRISPR/Cas9 Tool Kit for Efficient and Targeted Insertion/Deletion Mutagenesis of the Komagataella phaffii (Pichia pastoris) Genome",
abstract = "Efficient targeted genome engineering of Komagataella phaffii requires balanced expression of Cas9 nuclease and a target-specific guide RNA (gRNA). In addition, correct processing of the transcribed RNA to provide the designed gRNA as a target selective partner of targeted Cas9 protein for binding to genomic DNA is essential for efficient genome engineering. This method describes a step-by-step procedure and recommended tools for simple and efficient design of gRNAs to introduce insertions or deletions at targeted sites by CRISPR/Cas9-directed double-strand breaks, followed by error-prone nonhomologous end-joining repair.",
keywords = "CRISPR/Cas9, Genome engineering, INDELs, Komagataella phaffii, ORF interruption, Pichia pastoris",
author = "Fischer, {Jasmin Elgin} and Anton Glieder",
note = "Publisher Copyright: {\textcopyright} 2022, Springer Science+Business Media, LLC, part of Springer Nature.",
year = "2022",
doi = "10.1007/978-1-0716-2399-2_8",
language = "English",
isbn = "978-1-0716-2398-5",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "121--133",
editor = "V. Mapelli and Bettiga, {M. }",
booktitle = "Yeast Metabolic Engineering",
}