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A total of 48 h after transfection, cells were collected and RNA was extracted using the RNeasy Mini Kit (Qiagen, 74106), according to the manufacturers instructions

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Using a single CRISPR array encoding a pair of EMX1- and PVALB -targeting spacers, efficient cleavage at both loci was detected (Figure 4F, showing both a schematic design of the crRNA array and a Surveyor blot showing efficient mediation of cleavage)