Precise detection of a murine germline mutation of the Notch3 gene associated with kyphosis and developmental disorders
Humpback ( ) mice harbor a pathogenic mutation in the gene and can serve as a beneficial animal model for investigating human myopathy, kyphosis, and developmental disorders, including lateral meningocele syndrome. Detection of the point mutation in mice is important for maintaining strains and scru...
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Published in | Journal of advanced veterinary and animal research Vol. 8; no. 1; pp. 7 - 13 |
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Main Authors | , , , , , , |
Format | Journal Article |
Language | English |
Published |
Bangladesh
Network for the Veterinarians of Bangladesh Bangladesh Agricultural Universityת Faculty of Veterinary Science
01.03.2021
A periodical of the Network for the Veterinarians of Bangladesh (BDvetNET) Network for the Veterinarians of Bangladesh |
Subjects | |
Online Access | Get full text |
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Summary: | Humpback (
) mice harbor a pathogenic mutation in the
gene and can serve as a beneficial animal model for investigating human myopathy, kyphosis, and developmental disorders, including lateral meningocele syndrome. Detection of the point mutation in
mice is important for maintaining strains and scrutinizing genetic rescues, especially considering that homozygous mice are infertile and indistinguishable from their littermates at a young age. This study aimed for the development of a novel, precise, and time-saving genotyping method to identify the mutation in
mice.
In order to study the
mouse line, we describe how we applied several tools, including quantitative polymerase chain reaction (qPCR), multiplex tetra-primer amplification-refractory mutation system (ARMS-PCR) and Sanger sequencing, toward the recognition of heterozygous and homozygous mice.
The
mutation was clearly identified using qPCR and ARMS assays, but the latter was a more precise and cost-effective approach. The lengths of the ARMS-PCR amplicons are 210 bp and 164 bp for the wild-type and
alleles, respectively. Moreover, the genotyping results for each mouse were corroborated by Sanger DNA sequencing.
Our newly developed PCR-based ARMS system affords a swift and precise way to genotype the
mice. ARMS-PCR does not rely on any advanced equipment and is useful as a genotyping method for other model organisms that harbor a pathogenic variant. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 2311-7710 2311-7710 |
DOI: | 10.5455/javar.2021.h479 |