Generation and validation of a conditional knockout mouse model for the study of the Smith-Lemli-Opitz syndrome
Smith-Lemli-Opitz Syndrome (SLOS) is a developmental disorder (OMIM #270400) caused by autosomal recessive mutations in the Dhcr7 gene, which encodes the enzyme 3β-hydroxysterol-Δ7 reductase. SLOS patients present clinically with dysmorphology and neurological, behavioral, and cognitive defects, wit...
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| Format: | Article |
| Language: | English |
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Elsevier
2021-01-01
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| Series: | Journal of Lipid Research |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227520437046 |
| _version_ | 1828138548332593152 |
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| author | Babunageswararao Kanuri Vincent Fong Sithara Raju Ponny Keri A. Tallman Sriganesh Ramachandra Rao Ned Porter Steven J. Fliesler Shailendra B. Patel |
| author_facet | Babunageswararao Kanuri Vincent Fong Sithara Raju Ponny Keri A. Tallman Sriganesh Ramachandra Rao Ned Porter Steven J. Fliesler Shailendra B. Patel |
| author_sort | Babunageswararao Kanuri |
| collection | DOAJ |
| description | Smith-Lemli-Opitz Syndrome (SLOS) is a developmental disorder (OMIM #270400) caused by autosomal recessive mutations in the Dhcr7 gene, which encodes the enzyme 3β-hydroxysterol-Δ7 reductase. SLOS patients present clinically with dysmorphology and neurological, behavioral, and cognitive defects, with characteristically elevated levels of 7-dehydrocholesterol (7-DHC) in all bodily tissues and fluids. Previous mouse models of SLOS have been hampered by postnatal lethality when Dhcr7 is knocked out globally, while a hypomorphic mouse model showed improvement in the biochemical phenotype with aging and did not manifest most other characteristic features of SLOS. We report the generation of a conditional knockout of Dhcr7 (Dhcr7flx/flx), validated by generating a mouse with a liver-specific deletion (Dhcr7L-KO). Phenotypic characterization of liver-specific knockout mice revealed no significant changes in viability, fertility, growth curves, liver architecture, hepatic triglyceride secretion, or parameters of systemic glucose homeostasis. Furthermore, qPCR and RNA-Seq analyses of livers revealed no perturbations in pathways responsible for cholesterol synthesis, either in male or in female Dhcr7L-KO mice, suggesting that hepatic disruption of postsqualene cholesterol synthesis leads to minimal impact on sterol metabolism in the liver. This validated conditional Dhcr7 knockout model may now allow us to systematically explore the pathophysiology of SLOS, by allowing for temporal, cell and tissue-specific loss of DHCR7. |
| first_indexed | 2024-04-11T18:38:10Z |
| format | Article |
| id | doaj.art-fffd503a6747449ba298d727bfc30580 |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-04-11T18:38:10Z |
| publishDate | 2021-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-fffd503a6747449ba298d727bfc305802022-12-22T04:09:09ZengElsevierJournal of Lipid Research0022-22752021-01-0162100002Generation and validation of a conditional knockout mouse model for the study of the Smith-Lemli-Opitz syndromeBabunageswararao Kanuri0Vincent Fong1Sithara Raju Ponny2Keri A. Tallman3Sriganesh Ramachandra Rao4Ned Porter5Steven J. Fliesler6Shailendra B. Patel7Division of Endocrinology, Diabetes and Metabolism, University of Cincinnati, Cincinnati, OH, USADivision of Endocrinology, Diabetes and Metabolism, University of Cincinnati, Cincinnati, OH, USADivision of Human Genetics, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USADepartment of Chemistry and Vanderbilt Institute of Chemical Biology, Vanderbilt University, Nashville, TN, USADepartments of Ophthalmology and Biochemistry, Jacobs School of Medicine and Biomedical Sciences, University at Buffalo-State University of New York, Buffalo, NY, USADepartment of Chemistry and Vanderbilt Institute of Chemical Biology, Vanderbilt University, Nashville, TN, USADepartments of Ophthalmology and Biochemistry, Jacobs School of Medicine and Biomedical Sciences, University at Buffalo-State University of New York, Buffalo, NY, USA; Graduate Program in Neuroscience, University at Buffalo- State University of New York, Buffalo, NY, USA; Research Service, VA Western New York Healthcare System, Buffalo, NY, USADivision of Endocrinology, Diabetes and Metabolism, University of Cincinnati, Cincinnati, OH, USA; For correspondence: Shailendra B. Patel.Smith-Lemli-Opitz Syndrome (SLOS) is a developmental disorder (OMIM #270400) caused by autosomal recessive mutations in the Dhcr7 gene, which encodes the enzyme 3β-hydroxysterol-Δ7 reductase. SLOS patients present clinically with dysmorphology and neurological, behavioral, and cognitive defects, with characteristically elevated levels of 7-dehydrocholesterol (7-DHC) in all bodily tissues and fluids. Previous mouse models of SLOS have been hampered by postnatal lethality when Dhcr7 is knocked out globally, while a hypomorphic mouse model showed improvement in the biochemical phenotype with aging and did not manifest most other characteristic features of SLOS. We report the generation of a conditional knockout of Dhcr7 (Dhcr7flx/flx), validated by generating a mouse with a liver-specific deletion (Dhcr7L-KO). Phenotypic characterization of liver-specific knockout mice revealed no significant changes in viability, fertility, growth curves, liver architecture, hepatic triglyceride secretion, or parameters of systemic glucose homeostasis. Furthermore, qPCR and RNA-Seq analyses of livers revealed no perturbations in pathways responsible for cholesterol synthesis, either in male or in female Dhcr7L-KO mice, suggesting that hepatic disruption of postsqualene cholesterol synthesis leads to minimal impact on sterol metabolism in the liver. This validated conditional Dhcr7 knockout model may now allow us to systematically explore the pathophysiology of SLOS, by allowing for temporal, cell and tissue-specific loss of DHCR7.http://www.sciencedirect.com/science/article/pii/S0022227520437046dysmorphologycholesterol synthesis7-dehydrocholesterolSmith-Lemli-Opitz syndrome |
| spellingShingle | Babunageswararao Kanuri Vincent Fong Sithara Raju Ponny Keri A. Tallman Sriganesh Ramachandra Rao Ned Porter Steven J. Fliesler Shailendra B. Patel Generation and validation of a conditional knockout mouse model for the study of the Smith-Lemli-Opitz syndrome Journal of Lipid Research dysmorphology cholesterol synthesis 7-dehydrocholesterol Smith-Lemli-Opitz syndrome |
| title | Generation and validation of a conditional knockout mouse model for the study of the Smith-Lemli-Opitz syndrome |
| title_full | Generation and validation of a conditional knockout mouse model for the study of the Smith-Lemli-Opitz syndrome |
| title_fullStr | Generation and validation of a conditional knockout mouse model for the study of the Smith-Lemli-Opitz syndrome |
| title_full_unstemmed | Generation and validation of a conditional knockout mouse model for the study of the Smith-Lemli-Opitz syndrome |
| title_short | Generation and validation of a conditional knockout mouse model for the study of the Smith-Lemli-Opitz syndrome |
| title_sort | generation and validation of a conditional knockout mouse model for the study of the smith lemli opitz syndrome |
| topic | dysmorphology cholesterol synthesis 7-dehydrocholesterol Smith-Lemli-Opitz syndrome |
| url | http://www.sciencedirect.com/science/article/pii/S0022227520437046 |
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