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KCNQ1OT1 RT-PCR Real Time cod. BM-031 -Quantification assay of KCNQ1OT1
Principle of the test : Quantitative analysis of KCNQ1OT1 mRNA expression
Technology : Relatitive Quantitative Real Time PCR
Gene Target : KCNQ1OT1
Internal Control: GAPDH
Specimen : cDNA
Results : ΔΔCt method
Reporting Units : Arbitrary Units (AU)
Number of tests : 25 tests BM-031
Kit storage : -20°C
Necessary equipment : 7500 Real Time PCR System
Status: Ready to use
KCNQ1OT1 RT-PCR Real Time cod. BM-031
Quantitative Analysis of KCNQ1OT1 mRNA expression
Imprinting disorders are a group of congenital diseases caused by dysregulation of genomic imprinting, affecting prenatal and postnatal growth, neurocognitive development, metabolism and cancer predisposition. Aberrant expression of imprinted genes can be achieved through different mechanisms, classified into epigenetic - if not involving DNA sequence change - or genetic in the case of altered genomic sequence. Despite the underlying mechanism, the phenotype depends on the parental allele affected and opposite phenotypes may result depending on the involvement of the maternal or the paternal chromosome(1).
A large cluster of imprinted genes is located at chromosome 11p15.5. The cluster is divided into two independent domains, each controlled by a separate imprinting control regions (ICRs). The telomeric and centromeric ICRs (ICR1 and ICR2, respectively) work by different mechanisms. ICR2 is the promoter of the non-coding and imprinted KCNQ1OT1 gene. This is contained into and has antisense orientation with respect to the protein-coding gene KCNQ1. The long non-coding KCNQ1OT1 transcript silences in cis the imprinted genes of the centromeric domain on the paternal chromosome, although its mechanism of action is not fully understood2. On the maternal chromosome, ICR2 is methylated, KCNQ1OT1 is not transcribed and the flanking imprinted genes expressed(3).
Heterogeneous molecular defects affecting the 11p15.5 imprinted gene cluster are associated with the congenital growth disorders, Beckwith–Wiedemann syndrome (BWS) and Silver–Russell syndrome (SRS). The BWS (OMIM 130650) is characterized by overgrowth, macroglossia, abdominal wall defects and predisposition to embryonal tumors in childhood. The SRS (OMIM 180860) is associated with growth restriction, hypotonia and characteristic dysmorphic features. Opposite DNA methylation defects have been found at ICR1 in BWS and SRS and associated with reciprocal alterations of IGF2-H19 expression. Loss of the maternal-specific ICR2 methylation is the most frequent defect in BWS, resulting in the bi-allelic activation of KCNQ1OT1 and bi-allelic silencing of the centromeric domain genes, including the cell growth inhibitor CDKN1C. Mutations of CDKN1C account for 5% of the BWS cases(4).
Imprinting disorders are largely underdiagnosed because of the broad range of clinical signs, the overlap of presentation among different disorders, the presence of mild phenotypes, the mitigation of the phenotype with age and the limited availability of molecular techniques employed for diagnosis(1).
BioMole has developed and validated a novel quantitative analysis of KCNQ1OT1 mRNA expression. This method is based on real-time PCR. Relative quantification of mRNA expression of KCNQ1OT1 was achieved by normalization to the reference gene GAPDH. Relative quantification of target gene expression in patients was performed with the ΔΔCt method and the relative KCNQ1OT1 fold changes were determined. Results are expressed in corresponding arbitrary units (AU) User friendly and complete, the KCNQ1OT1 RT-PCR Real Time kit is suitable for any laboratory.
Reference
1. Syndromic Disorders Caused by Disturbed Human Imprinting
Diana Carli, Evelise Riberi, Giovanni Battista Ferrero, and Alessandro Mussa.
J.Clin Res Pediatr Endocrinol 2020;12(1):1-16.
2. Kcnq1ot1: a chromatin regulatory RNA.
Kanduri C. Semin. Cell Dev. Biol. 2011 February 21
3. The KCNQ1OT1 imprinting control region and non-coding RNA: new properties derived from the study of Beckwith–Wiedemann syndrome and Silver–Russell syndrome cases
Nicoletta Chiesa, Agostina De Crescenzo, Kankadeb Mishra, Lucia Perone, Massimo Carella, Orazio Palumbo, Alessandro Mussa, Angela Sparago, Flavia Cerrato, Silvia Russo, Elisabetta Lapi, Maria Vittoria Cubellis, Chandrasekhar Kanduri, Margherita Cirillo Silengo, Andrea Riccio, and Giovanni Battista Ferrero
Hum Mol Genet. 2012 Jan 1; 21(1): 10–25
4. Beckwith—Wiedemann syndrome
Choufani S., Shuman C., Weksberg R.. Am. J. Med. Genet. C Semin. Med. Genet. 2010.