Genome editing in penicillium chrysogenum using cas9 ribonucleoprotein particles

Carsten Pohl; László Mózsik; Arnold J.M. Driessen; Roel A.L. Bovenberg; Yvonne I. Nygård

doi:10.1007/978-1-4939-7795-6_12

Genome editing in penicillium chrysogenum using cas9 ribonucleoprotein particles

Carsten Pohl, László Mózsik, Arnold J.M. Driessen^*, Roel A.L. Bovenberg, Yvonne I. Nygård

^*Corresponding author for this work

Not published at VTT

Research output: Chapter in Book/Report/Conference proceeding › Chapter or book article › Scientific › peer-review

19 Citations (Scopus)

Abstract

Several CRISPR/Cas9 tools have been recently established for precise genome editing in a wide range of filamentous fungi. This genome editing platform offers high flexibility in target selection and the possibility of introducing genetic deletions without the introduction of transgenic sequences. This chapter describes an approach for the transformation of Penicillium chrysogenum protoplasts with preassembled ribonucleoprotein particles (RNPs) consisting of purified Cas9 protein and in vitro transcribed single guide RNA (sgRNA) for the deletion of genome sequences or their replacement with alternative sequences. This method is potentially transferable to all fungal strains where protoplasts can be obtained from.

Original language	English
Title of host publication	Synthetic Biology: Methods and Protocols
Publisher	Humana Press
Pages	213-232
ISBN (Electronic)	978-1-4939-7795-6
ISBN (Print)	978-1-4939-7794-9, 978-1-4939-9287-4
DOIs	https://doi.org/10.1007/978-1-4939-7795-6_12
Publication status	Published - 2018
MoE publication type	A3 Part of a book or another research book

Publication series

Series	Methods in Molecular Biology
Volume	1772
ISSN	1064-3745

Funding

The work was supported by the People Programme (Marie Curie Actions) of the European Union’s Seventh Framework Programme FP7/2007-2013/under REA grant agreement no. [607332], and the Marie Skłodowska-Curie Co-funding of regional, national and international programmes (COFUND-DP) ALERT programme under REA grant agreement no. [713482].

Keywords

CRISPR/Cas9
Filamentous fungi
Genome editing
HDR
NHEJ
Penicillium chrysogenum
Ribonucleoprotein particle
RNP
Endonucleases/genetics
Protoplasts/metabolism
Gene Targeting/methods
Gene Editing/methods
RNA, Guide, CRISPR-Cas Systems/genetics
CRISPR-Cas Systems/genetics
Clustered Regularly Interspaced Short Palindromic Repeats/genetics
Gene Deletion
Ribonucleoproteins/genetics
Penicillium chrysogenum/genetics

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10.1007/978-1-4939-7795-6_12

Cite this

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abstract = "Several CRISPR/Cas9 tools have been recently established for precise genome editing in a wide range of filamentous fungi. This genome editing platform offers high flexibility in target selection and the possibility of introducing genetic deletions without the introduction of transgenic sequences. This chapter describes an approach for the transformation of Penicillium chrysogenum protoplasts with preassembled ribonucleoprotein particles (RNPs) consisting of purified Cas9 protein and in vitro transcribed single guide RNA (sgRNA) for the deletion of genome sequences or their replacement with alternative sequences. This method is potentially transferable to all fungal strains where protoplasts can be obtained from.",

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Genome editing in penicillium chrysogenum using cas9 ribonucleoprotein particles. / Pohl, Carsten; Mózsik, László; Driessen, Arnold J.M. et al.
Synthetic Biology: Methods and Protocols. Humana Press, 2018. p. 213-232 (Methods in Molecular Biology, Vol. 1772).

Research output: Chapter in Book/Report/Conference proceeding › Chapter or book article › Scientific › peer-review

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KW - Gene Targeting/methods

KW - Gene Editing/methods

KW - RNA, Guide, CRISPR-Cas Systems/genetics

KW - CRISPR-Cas Systems/genetics

KW - Clustered Regularly Interspaced Short Palindromic Repeats/genetics

KW - Gene Deletion

KW - Ribonucleoproteins/genetics

KW - Penicillium chrysogenum/genetics

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ER -

Genome editing in penicillium chrysogenum using cas9 ribonucleoprotein particles

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