Strategies for genetic inactivation of long noncoding RNAs in zebrafish

Effective inactivation of the lncRNA malat1 in zebrafish by insertion of a premature polyadenylation signal. (A) The malat1 locus in zebrafish. Shown are the corresponding CAGE (Nepal et al. 2013; Haberle et al. 2014), H3K4me3 ChIP-Seq (Ulitsky et al. 2011), RNA-seq, and H3K27ac ChIP-Seq (Pérez-Rico et al. 2017) tracks from WT zebrafish. (B) Generation of the malat1^poly(A) allele by targeted knock-in of the poly(A) signal. The hybridization site of the RNA blot probe is indicated as a gray box. ha, homology arms. (C) malat1 expression in WT and homozygous malat1^poly(A) embryos detected by an RNA blot. 18S rRNA was used as a loading reference. hpf, hours postfertilization. (D) malat1 expression across WT and homozygous malat1^poly(A) adult zebrafish tissues detected by RNA blot. 18S rRNA was used as a loading reference.