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. 2019 Nov 4;10:1246. doi: 10.3389/fphar.2019.01246

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Copyright © 2019 Filippova, Matveeva, Zhuravlev, Balakhonova, Prokhorova, Malanin, Shah Mahmud, Grigoryeva, Anufrieva, Semenov, Vlassov and Stepanov

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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(A–C) Design of sgRNAs targeted at Gas5 box C/D snoRNAs. (A)—Exon/intron structure of Gas5 with positions of SNORDs. (B)—Sequences of the selected target snoRNAs with positions of PAM sequences (underlined) and CRISPR/Cas9 cleavage sites (asterisks). C and D boxes are indicated by green and purple font, respectively. Nucleotide complementary to the 2’-O-methylation site in the target rRNA is indicated in bold red. (C)—Sequences of the top and bottom strands of sgRNAs. Overhangs for ligation into the pair of BstV2I sites in pX458 are shown in blue italics.