KIT FOR THE GENOTYPING OF MICE hyh MUTANTS FOR PROTEIN a-Snap
Description
The mutant mouse hyh (hydrocephalus with hop gait) develops congenital hydrocephalus due to an autosomal recessive mutation located on chromosome 7. The mutation presented by the hyh mice corresponds to a point mutation in the a-Snap gene (soluble alpha N- ethylmaleimide sensitive factor attachment protein).
The a-Snap protein is constitutively expressed in all cells of the body and plays a very important role in vesicle-mediated intercompartmental protein trafficking. The alteration of the vesicular traffic towards the plasma membrane affects the transport of adhesion proteins and / or proteins of the same cell membrane. Until now, the interest of this strain of mutant mice has been focused exclusively on research on neuropathological alterations associated with congenital hydrocephalus, since it is a good animal model to study the etiology of this disease in humans, but currently , some authors propose a-Snap as a future therapeutic target in diseases such as diabetes, cancer and diseases of the central nervous system.
Thus, the present invention relates to a method for genotyping mutant hyh mice for the a-Snap protein as well as a genotyping kit to carry out said method and oligonucleotides specifically designed to be used in said method.
Advantages
Until 2004, animals homozygous for the mutated gene could only be distinguished phenotypically and histologically. Since 2004, with the identification of the mutated gene, the identification of homozygous animals has been possible only through PCR amplification and sequencing of the fragment that contains the sequence. Furthermore, it was not possible to genotype hyh mice by PCR-RFLP due to the absence of a restriction enzyme whose target allows the wild allele to be differentiated from the mutated allele. With the aim of genotyping these mice, the present invention provides primers that not only make it possible to amplify a sequence that includes the locus of the hyh mutation but are also capable of creating a new restriction site in the amplified fragment from the snap + allele, but not in the amplified from snap-, which allows distinguishing between the different genotypes (snap + / +; snap +/-; snap - / -) in a simple, reliable and inexpensive way.
Uses and Applications
The present technology has its usefulness to perform, in a reliable and economic way, the genotyping of mutant hyh mice for the a-Snap protein, with the aim of continuing to advance in the medical areas related to congenital hydrocephalus or other diseases related to such protein.
Keywords
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Areas
Applicants
UNIVERSIDAD DE MÁLAGA, UNIVERSIDAD AUSTRAL DE CHILE, UNIVERSIDAD AUSTRAL DE CHILE
Inventors
PATRICIA PÁEZ GONZÁLEZ, ANTONIO JESUS JIMENEZ LARA, RUTH ROALES BUJÁN, ISABEL MATAS CASADO, LUIS FEDERICO BÁTIZ , JOSE MANUEL FERNANDEZ, LUIS MANUEL RODRÍGUEZ PÉREZ, CAYO JUAN RAMOS RODRIGUEZ, ESTEBAN MARTÍN RODRÍGUEZ CAIRO
Filing Date
31/12/2008
Protection Level: National (Spain)
Processing Status: Spanish patent
