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Generation of the Sotos Syndrome Deletion in Mice

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Generation of the Sotos Syndrome Deletion in Mice Edinburgh Research Explorer Generation of the Sotos syndrome deletion in mice Citation for published version: Sanger Mouse Genetics Project, Migdalska, AM, van der Weyden, L, Ismail, O, Rust, AG, Rashid, M, White, JK, Sánchez-Andrade, G, Lupski, JR, Logan, DW, Arends, MJ & Adams, DJ 2012, 'Generation of the Sotos syndrome deletion in mice', Mammalian Genome, vol. 23, no. 11-12, pp. 749-57. https://doi.org/10.1007/s00335-012-9416-0 Digital Object Identifier (DOI): 10.1007/s00335-012-9416-0 Link: Link to publication record in Edinburgh Research Explorer Document Version: Publisher's PDF, also known as Version of record Published In: Mammalian Genome Publisher Rights Statement: Open Access This article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited. General rights Copyright for the publications made accessible via the Edinburgh Research Explorer is retained by the author(s) and / or other copyright owners and it is a condition of accessing these publications that users recognise and abide by the legal requirements associated with these rights. Take down policy The University of Edinburgh has made every reasonable effort to ensure that Edinburgh Research Explorer content complies with UK legislation. If you believe that the public display of this file breaches copyright please contact [email protected] providing details, and we will remove access to the work immediately and investigate your claim. Download date: 05. Oct. 2021 Mamm Genome (2012) 23:749–757 DOI 10.1007/s00335-012-9416-0 Generation of the Sotos syndrome deletion in mice Anna M. Migdalska • Louise van der Weyden • Ozama Ismail • The Sanger Mouse Genetics Project • Alistair G. Rust • Mamunur Rashid • Jacqueline K. White • Gabriela Sa´nchez-Andrade • James R. Lupski • Darren W. Logan • Mark J. Arends • David J. Adams Received: 11 June 2012 / Accepted: 16 July 2012 / Published online: 29 August 2012 Ó The Author(s) 2012. This article is published with open access at Springerlink.com Abstract Haploinsufficiency of the human 5q35 region for their gestational age and also showed decreased spanning the NSD1 gene results in a rare genomic disorder postnatal growth, in contrast to Sotos patients. known as Sotos syndrome (Sotos), with patients displaying Df(13)Ms2Dja?/- mice did, however, display deficits in a variety of clinical features, including pre- and postnatal long-term memory retention and dilation of the pelvicaly- overgrowth, intellectual disability, and urinary/renal ceal system, which in part may model the learning diffi- abnormalities. We used chromosome engineering to gen- culties and renal abnormalities observed in Sotos patients. erate a segmental monosomy, i.e., mice carrying a het- Thus, haploinsufficiency of genes within the mouse erozygous 1.5-Mb deletion of 36 genes on mouse 4732471D19Rik–B4galt7 deletion interval play important chromosome 13 (4732471D19Rik-B4galt7), syntenic with roles in growth, memory retention, and the development of 5q35.2–q35.3 in humans (Df(13)Ms2Dja?/- mice). Sur- the renal pelvicalyceal system. prisingly Df(13)Ms2Dja?/- mice were significantly smaller Introduction Electronic supplementary material The online version of this article (doi:10.1007/s00335-012-9416-0) contains supplementary material, which is available to authorized users. Sotos syndrome (Sotos; MIM# 117550) is an autosomal dominant, multiple-anomaly syndrome characterized by A. M. Migdalska Á L. van der Weyden Á overgrowth, a distinctive craniofacial appearance, A. G. Rust Á M. Rashid Á D. J. Adams (&) Experimental Cancer Genetics, Wellcome Trust Sanger Institute, advanced bone age, and variable learning disabilities. Wellcome Trust Genome Campus, Hinxton, However, there is significant clinical heterogeneity in Sotos Cambridge CB10 1HH, UK syndrome, with some affected individuals also showing e-mail: [email protected] frequent ear and chest infections, cardiac and urinary/renal O. Ismail Á The Sanger Mouse Genetics Project Á J. K. White defects, seizures, scoliosis, and behavioural problems Mouse Genetics Project, Wellcome Trust Sanger Institute, (Tatton-Brown et al. 1993). The diagnosis of Sotos syn- Wellcome Trust Genome Campus, Hinxton, drome relied solely on clinical criteria until haploinsuffi- Cambridge CB10 1HH, UK ciency of the NSD1 gene (encoding a histone G. Sa´nchez-Andrade Á D. W. Logan methyltransferase implicated in chromatin regulation) was Genetics of Instinctive Behaviour, Wellcome Trust Sanger identified as causative (Kurotaki et al. 2002). Subsequent Institute, Wellcome Trust Genome Campus, Hinxton, analysis of patients clinically diagnosed with Sotos showed Cambridge CB10 1HH, UK that haploinsufficiency of NSD1 due to intragenic NSD1 J. R. Lupski mutations, partial NSD1 deletions, or chromosomal mic- Department of Molecular and Human Genetics, Baylor College rodeletions spanning the 5q35 region encompassing the of Medicine, Houston, TX 77030, USA entire NSD1 gene accounted for more than 90 % of cases (with the prevalence of intragenic NSD1 mutations and M. J. Arends Department of Pathology, Addenbrooke’s Hospital, University 5q35 microdeletions encompassing the NSD1 gene of Cambridge, Cambridge CB2 0QQ, UK depending greatly on ethnic origin) (Baujat et al. 2004; 123 750 A. M. Migdalska et al.: Sotos syndrome deletion in mice Tatton-Brown and Rahman 2007; Tatton-Brown et al. Materials and methods 2005a, b). It has also been proposed that GPC3 mutations or 11p15 abnormalities or both may be responsible for some Gene targeting in ES cells and generation of deletion Sotos cases without NSD1 abnormalities (Baujat et al. 2004; mice Li et al. 2001; Tatton-Brown and Rahman 2007). The major clinical features of Sotos, including over- The 50 Hprt MICER targeting vector MHPN55m07 (Adams growth, facial abnormalities, and intellectual disabilities, et al. 2004) was linearized with KpnI and electroporated into are diagnosed in Sotos patients with intragenic NSD1 E14Tg2a embryonic stem (ES) cells (129P2Ola), which were mutations and in Sotos individuals carrying 5q35 mic- selected in G418 as described previously (Ramirez-Solis et al. rodeletions (Tatton-Brown et al. 2005a, b). However, in 1995). Southern blotting was performed on BstEII-digested ES contrast to Sotos individuals with NSD1 mutations, Sotos cell genomic DNA (gDNA) using a probe amplified from patients with 5q35 microdeletions tend to show less pro- E14Tg2a gDNA (50-GTC TGT TGT TAA AAG CTA AAA nounced overgrowth but more profound intellectual dis- CCT TAG A-30 and 50-TGA GCT ACA GTT TGG TTC TGG ability (Kurotaki et al. 2003; Nagai et al. 2003; Saugier- TGG ATA AAC-30) to identify correctly targeted clones. The Veber et al. 2007; Tatton-Brown et al. 2005a), and several 30 Hprt MICER targeting vector MHPP265c24 (Adams et al. studies have reported an increased frequency of cardio- 2004) was linearized with NcoI and electroporated into vascular and urinary/renal abnormalities in 5q35 microde- MHPN55m07-targeted E14Tg2a ES cells, which were then letion Sotos patients (Kurotaki et al. 2003; Nagai et al. selected in puromycin. Southern blotting was performed on 2003; Saugier-Veber et al. 2007). Thus, it is possible that SpeI-digested gDNA using a probe amplified from E14Tg2a genes other than the NSD1 gene could be dosage-sensitive gDNA (50-CAG TAA TAT AGT AGA AGC ATG GTC CAT- and therefore responsible for the extended variability and 30 and 50-ATG ATA CTG AAC ACA GAC AAC AGA GGC degree of severity of phenotypes observed in Sotos patients TGC T-30). Double-targeted ES cell clones were electropora- who carry 5q35 microdeletions. Furthermore, selected ted with a Cre-expression vector and selected in hypoxanthine, patients with Sotos can have low laboratory values of the aminopterin, and thymidine (HAT) medium, as described factor XII blood-clotting protein, a phenotype thought to previously (Ramirez-Solis et al. 1995), to identify whether the result from the hemizygous deletion of 5q35, unmasking a 50 Hprt and 30 Hprt vectors were targeted in cis or in trans. functional single nucleotide variant (SNV) on the remaining Clones of ES cells that had undergone cis recombination were nondeleted allele of the FXII gene within the common Sotos identified and confirmed by PCR (50-AAG GGT GTT TAT deletion interval (Kurotaki et al. 2005). The latter genetic TCC CCA TGG ACT AAT TAT G-30 and 50-CCT TCA TCA mechanism has recently also been shown to be responsible CAT CTC GAG CAA GAC GTT CAG-30; presence of a 1.7-kb for TAR (thrombocytopenia absent radius) syndrome where band confirmed the cis orientation). The deletion allele was 1q21.1 deletions appear to unmask functional variants designated Df(13)Ms2Dja. ES cell clones carrying the con- associated with the RBM8a gene (Albers et al. 2012). ditional deletion (prior to electroporation with Cre) were The mouse orthologue of the NSD1 gene (Nsd1) is highly injected into C57BL/6-Tyrc-Brd blastocysts and transmitted conserved (83 % homology at amino acid level) (Kurotaki through the germline. F1 mice carrying the conditional dele- et al. 2001); however, heterozygous Nsd1 mice do not dis- tion were bred with CMV-Cre mice (Su et al. 2002) to gen- play any gross phenotypic abnormalities (Rayasam et al. erate heterozygous Df(13)Ms2Dja mice (Df(13)Ms2Dja?/-). 2003). Similarly, mice carrying knockout mutations of other These deficiency mice, Df(13)Ms2Dja?/-, were backcrossed genes within the most frequently detected human Sotos 5q35 to C57BL/6J for three to four generations and maintained on microdeletion interval have failed to display dominant phe- a mixed C57BL/6J-129P2Ola background. All mice were notypes, suggesting that Sncb, Unc5a, Fgfr4, Mxd3, Rgs14, housed and experimental procedures were carried out in Slc34a1, F12, Grk6, Dbn1, and Dok3 are not dosage-sensi- accordance with UK Home Office guidelines. tive and, therefore, at least individually are not likely to be responsible for the extended variability and degree of Comparative genomic hybridization (CGH) severity of clinical features observed in 5q35 microdeletion Sotos patients (Mouse Genome Informatics; http://www.
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