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. 2019 Jul 1;33(13-14):741–746. doi: 10.1101/gad.326363.119

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© 2019 Vitali and Kiss; Published by Cold Spring Harbor Laboratory Press

This article, published in Genes & Development, is available under a Creative Commons License (Attribution 4.0 International), as described at http://creativecommons.org/licenses/by/4.0/.

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Figure 2. — Construction of SNORD97 knockout and SCARNA97 knockout HAP1 cells. (A) Strategy for CRISPR–Cas9-mediated excision of SNORD97 and SCARNA97 genes. The intronic SNORD97 and SCARNA97 genes (open arrows), the neighboring exons of the EIF4G2 (E13 and E14) and LARP4 (E10 and E11) host genes, and the forward (F1 and F2) and reverse (R1 and R2) primers used for PCR analyses are shown. The genomic sequences targeted by sgRNAs are indicated in sense orientation. The PAM (or complementary) sequences are in italics. The D boxes are underlined. (B) PCR analyses of genomic DNAs obtained from SNORD97 knockout (SNO97-KO) and SCARNA97 knockout (SCA97-KO) cells. The amplified DNAs were analyzed on 2% agarose gels. Structures and expected sizes of the amplified genomic fragments are shown. (Lane M) DNA size marker in base pairs.