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. 2020 May 26;21(11):3747. doi: 10.3390/ijms21113747

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Nuclear localization of RNP and ssODN. (A) Experimental design. (B) RNP (HA-conjugated SpCas9 and sgRNAs targeting Rosa26 locus) and Cy-3-conjugated 100 bp-sized ssODN were electroporated into embryos at 25 h and 29 h after hCG injection. Next, half were immunostained with HA-Alexa 488 (green) mAb, and the other half were stained after two hours with the same target. Representative images are shown. The related videos are presented in Videos 1 and 2, Supplementary Materials. White solid line: intact nuclear membrane; white dotted line: disappeared nuclear membrane. (C) Physical barriers outside the chromosome (white arrow); blue: chromosome; red: ssODN; green: RNP.