4.5SH RNA counteracts deleterious exonization of SINE B1 in mice

4.5SH RNA is a highly abundant, small rodent-specific noncoding RNA that localizes to nuclear speckles en-riched in pre-mRNA-splicing regulators. To investigate the physiological functions of 4.5SH RNA, we have created mutant mice that lack the expression of 4.5SH RNA. The mutant mice exhibited embryonic lethality, suggesting that 4.5SH RNA is an essential species-specific noncoding RNA in mice. RNA-sequencing ana-lyses revealed that 4.5SH RNA protects the transcriptome from abnormal exonizations of the antisense inser-tions of the retrotransposon SINE B1 (asB1), which would otherwise introduce deleterious premature stop codons or frameshift mutations. Mechanistically, 4.5SH RNA base pairs with complementary asB1-contain-ing exons via the target recognition region and recruits effector proteins including Hnrnpm via its 50 stem loop region. The modular organization of 4.5SH RNA allows us to engineer a programmable splicing regulator to induce the skipping of target exons of interest. Our results also suggest the general existence of splicing reg-ulatory noncoding RNAs.