Gene Excision by Dual-Guide CRISPR-Cas9

Michael Spagnuolo; Mark Blenner

doi:10.1007/978-1-0716-1414-3_5

Gene Excision by Dual-Guide CRISPR-Cas9

Methods Mol Biol. 2021:2307:85-94. doi: 10.1007/978-1-0716-1414-3_5.

Authors

Michael Spagnuolo¹, Mark Blenner^{2

3}

Affiliations

¹ Department of Chemical and Biomolecular Engineering, Clemson University, Clemson, SC, USA.
² Department of Chemical and Biomolecular Engineering, Clemson University, Clemson, SC, USA. blenner@udel.edu.
³ Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, DE, USA. blenner@udel.edu.

PMID: 33847983
DOI: 10.1007/978-1-0716-1414-3_5

Abstract

CRISPR-Cas9 is frequently used for creating double-strand DNA breaks that result in indels through non-homologous end joining. Indels can revert to wild-type sequence and require sequencing or complex assays to measure. Cutting by two guide RNAs can lead to single indels at either cut site or simultaneous cutting at both sites and repair leading to gene excision.

Keywords: CRISPR-Cas9; Gene excision; Genome editing; Metabolic engineering; Synthetic biology; Yarrowia lipolytica.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

CRISPR-Cas Systems
Gene Editing / methods*
Genes, Fungal
Metabolic Engineering
RNA, Guide, CRISPR-Cas Systems / genetics*
Synthetic Biology
Yarrowia / genetics*

Substances

RNA, Guide, CRISPR-Cas Systems