Delf-Magnus Kummerfeld

Prader–Willi syndrome (PWS) is a neurodevelopmental disorder characterized by distinctive developmental transition from neonatal hypotonia and poor feeding to progressive weight gain and, later, hyperphagia and obesity. The paternally expressed SNORD116 cluster is critical for postnatal growth, but how its deficiency impacts the progression of PWS remains unclear. Here, we analyzed the day-over-day growth rates for 351 mice with Snord116 deletion or altered Snord116 dosage, including their wild-type littermates, across both sexes and two genetic backgrounds. Snord116 expression increased markedly during early brain development. The Snord116-deficient mice exhibited a distinct three-stage growth trajectory: impaired growth until approximately P16, normalization between P17 and P21, followed by transient accelerated weight gain until approximately P30. Although growth rates subsequently normalized, the accumulated deficit was not fully compensated, and the adult Snord116KO mice remained smaller than the wild-type controls. These developmental transitions broadly parallel the early nutritional phases of human PWS, although the mice did not progress to exhibit persistent obesity-associated growth. Reactivation of the maternal Snord116 cluster partially rescued the growth phenotype, whereas activation of both parental alleles produced a modest growth advantage, which was most evident in the males. These findings identify early postnatal development as a critical window of Snord116-dependent growth regulation and provide a framework for investigating the mechanisms underlying PWS’s progression.

Prader–Willi syndrome (PWS) is a neurodevelopmental disorder characterized by distinctive developmental transition from neonatal hypotonia and poor feeding to pro-gressive weight gain and, later, hyperphagia and obesity. The paternally expressed SNORD116 cluster is critical for postnatal growth, but how its deficiency shapes this progression remains unclear. Here, we analyzed day-over-day growth rates in 351 mice carrying Snord116 deletion or altered Snord116 dosage, including wild-type littermates, across both sexes and two genetic backgrounds. Snord116 expression increased markedly during early brain development. Snord116-deficient mice exhibited a distinct three-stage growth trajectory: impaired growth until approximately P16, normalization between P17 and P21, followed by transient accelerated weight gain until approximately P30. Alt-hough growth rates subsequently normalized, the accumulated deficit was not fully compensated, and adult Snord116KO mice remained smaller than wild-type controls. These developmental transitions broadly parallel early nutritional phases of human PWS, although mice did not progress to persistent obesity-associated growth. Reactivation of the maternal Snord116 cluster partially rescued the growth phenotype, whereas activation of both parental alleles produced a modest growth advantage, most evident in males. These findings identify early postnatal development as a critical window of Snord116-dependent growth regulation and provide a framework for investigating mechanisms underlying PWS progression.

Prader–Willi syndrome (PWS) is a complex neurodevelopmental disorder caused by the deletion or inactivation of paternally expressed imprinted genes at the chromosomal region 15q11–q13. The PWS-critical region (PWScr) harbors tandemly repeated non-protein coding IPW-A exons hosting the intronic SNORD116 snoRNA gene array that is predominantly expressed in brain. Paternal deletion of PWScr is associated with key PWS symptoms in humans and growth retardation in mice (PWScr model). Dysregulation of the hypothalamic–pituitary axis (HPA) is thought to be causally involved in the PWS phenotype. Here we performed a comprehensive reverse transcription quantitative PCR (RT-qPCR) analysis across nine different brain regions of wild-type (WT) and PWScr mice to identify stably expressed reference genes. Four methods (Delta Ct, BestKeeper, Normfinder and Genorm) were applied to rank 11 selected reference gene candidates according to their expression stability. The resulting panel consists of the top three most stably expressed genes suitable for gene-expression profiling and comparative transcriptome analysis of WT and/or PWScr mouse brain regions. Using these reference genes, we revealed significant differences in the expression patterns of Igfbp7, Nlgn3 and three HPA associated genes: Pcsk1, Pcsk2 and Nhlh2 across investigated brain regions of wild-type and PWScr mice. Our results raise a reasonable doubt on the involvement of the Snord116 in posttranscriptional regulation of Nlgn3 and Nhlh2 genes. We provide a valuable tool for expression analysis of specific genes across different areas of the mouse brain and for comparative investigation of PWScr mouse models to discover and verify different regulatory pathways affecting this complex disorder.

Prader-Willi syndrome (PWS) is a neurogenetic multifactorial disorder caused by the deletion or inactivation of paternally imprinted genes on human chromosome 15q11-q13. The affected homologous locus is on mouse chromosome 7C. The positional conservation and organization of genes including the imprinting pattern between mice and men implies similar physiological functions of this locus. Therefore, considerable efforts to recreate the pathogenesis of PWS have been accomplished in mouse models. We provide a summary of different mouse models that were generated for the analysis of PWS and discuss their impact on our current understanding of corresponding genes, their putative functions and the pathogenesis of PWS. Murine models of PWS unveiled the contribution of each affected gene to this multi-facetted disease, and also enabled the establishment of the minimal critical genomic region (PWScr) responsible for core symptoms, highlighting the importance of non-protein coding genes in the PWS locus. Although the underlying disease-causing mechanisms of PWS remain widely unresolved and existing mouse models do not fully capture the entire spectrum of the human PWS disorder, continuous improvements of genetically engineered mouse models have proven to be very powerful and valuable tools in PWS research.

Knock-in genome targeting risks: Comprehensive locus analysis is essential for precision chromosome-editing identification.

Serotonin 5-HT2C receptor is a G-protein coupled excitatory receptor that regulates several biochemical pathways and has been implicated in obesity, mental state, sleep cycles, autism, neuropsychiatric disorders and neurodegenerative diseases. The activity of 5-HT2CR is regulatedviaalternative splicing and A to I editing of exon Vb of its pre-mRNA. Snord115 is a small nucleolar RNA that is expressed in mouse neurons and displays an 18-nucleotide base complementary to exon Vb of 5-HT2CR pre-mRNA. For almost two decades this putative guide element of Snord115 has wandered like a ghost through the literature in attempts to elucidate the biological significance of this complementarity. In mice, Snord115 is expressed in neurons and absent in the choroid plexus where, in contrast, 5-Ht2cr mRNA is highly abundant. Here we report the analysis of 5-Ht2cr pre-mRNA posttranscriptional processingviaRNA deep sequencing in a mouse model that ectopically expresses Snord115 in the choroid plexus. In contrast to previous reports, our analysis demonstrated that Snord115 does not control alternative splicing of 5-Ht2cr pre-mRNAin vivo. We identified a modest, yet statistically significant reduction of 5-Ht2cr pre-mRNA A to I editing at the major A, B, C and D sites. We suggest that Snord115 and exon Vb of 5Ht2cr pre-mRNA form a double-stranded structure that is subject to ADAR-mediated A to I editing. To the best of our knowledge, this is the first comprehensive Snord115 gain-of-function analysis based onin vivomouse models.