Universität zu Köln

Troeder, Simon E., Ebert, Lena K., Butt, Linus, Assenmacher, Sonja, Schermer, Bernhard ORCID: 0000-0002-5194-9000 and Zevnik, Branko ORCID: 0000-0001-7845-9522 (2018). An optimized electroporation approach for efficient CRISPR/Cas9 genome editing in murine zygotes. PLoS One, 13 (5). SAN FRANCISCO: PUBLIC LIBRARY SCIENCE. ISSN 1932-6203

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Abstract

Electroporation of zygotes represents a rapid alternative to the elaborate pronuclear injection procedure for CRISPR/Cas9-mediated genome editing in mice. However, current protocols for electroporation either require the investment in specialized electroporators or corrosive pre-treatment of zygotes which compromises embryo viability. Here, we describe an easily adaptable approach for the introduction of specific mutations in C57BL/6 mice by electroporation of intact zygotes using a common electroporator with synthetic CRISPR/ Cas9 components and minimal technical requirement. Direct comparison to conventional pronuclear injection demonstrates significantly reduced physical damage and thus improved embryo development with successful genome editing in up to 100% of living offspring. Hence, our novel approach for Easy Electroporation of Zygotes (EEZy) allows highly efficient generation of CRISPR/Cas9 transgenic mice while reducing the numbers of animals required.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Troeder, Simon E. UNSPECIFIED UNSPECIFIED UNSPECIFIED Ebert, Lena K. UNSPECIFIED UNSPECIFIED UNSPECIFIED Butt, Linus UNSPECIFIED UNSPECIFIED UNSPECIFIED Assenmacher, Sonja UNSPECIFIED UNSPECIFIED UNSPECIFIED Schermer, Bernhard UNSPECIFIED orcid.org/0000-0002-5194-9000 UNSPECIFIED Zevnik, Branko UNSPECIFIED orcid.org/0000-0001-7845-9522 UNSPECIFIED
URN:	urn:nbn:de:hbz:38-186694
DOI:	10.1371/journal.pone.0196891
Journal or Publication Title:	PLoS One
Volume:	13
Number:	5
Date:	2018
Publisher:	PUBLIC LIBRARY SCIENCE
Place of Publication:	SAN FRANCISCO
ISSN:	1932-6203
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language ONE-STEP GENERATION; MOUSE ZYGOTES; ZONA-PELLUCIDA; MICE; DELIVERY; RIBONUCLEOPROTEINS; KNOCKOUT; PROTEIN; EMBRYOS; CAS9 Multiple languages Multidisciplinary Sciences Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/18669