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. 2017 Sep;23(9):1385–1392. doi: 10.1261/rna.060723.117

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© 2017 Silva et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society

This article, published in RNA, 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 4. — Nef-sp knockout mice are viable and fertile. (A) Gene organization of mouse Nef-sp and mNEF-sp protein domain architecture. (B) Guide RNAs (gRNAs) used for generation of a 23-bp deletion in the mouse Nef-sp gene locus of single-cell embryos. (C) Premature translation stop codon (D220*) introduced by the deletion leading to a truncation of the protein before the nuclease domain. (D) Genotyping of the wild-type, Nef-sp^+/− and Nef-sp^−/− mice. Cartoon showing the set of primers (represented by arrows) used to distinguish the wild-type allele from the knockout allele. See Materials and Methods for details. (E) Representative mouse testes from wild-type and mutant Nef-sp animals showing normal testes size. Mutant mice show normal fertility. (F) Analysis of the total testicular transcriptome from heterozygous and homozygous Nef-sp knockout mutant mice. Overall genome annotations of reads do not show any changes. (G) Comparison of read counts of sense-oriented gene exonic and intronic reads reveal down-regulation of Nef-sp transcript, as expected. No other significant change in transcripts is noted. (H) Analysis of sense-oriented repeat reads shows an up-regulation of the indicated repeat elements. Table showing fold-changes in read counts of sense-oriented repeat elements and Nef-sp transcript.