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A fast and reliable method for detecting SNP rs67384697 (Hsa‐miR‐148a binding site) by a single run of allele‐specific real‐time PCR
Author(s) -
Malnati Mauro S.,
Biswas Priscilla,
Ugolotti Elisabetta,
Di Marco Eddi,
Sironi Francesca,
Parolini Francesca,
Garbarino Lucia,
Mazzocco Michela,
Zipeto Donato,
Biassoni Roberto
Publication year - 2020
Publication title -
hla
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.347
H-Index - 99
eISSN - 2059-2310
pISSN - 2059-2302
DOI - 10.1111/tan.13971
Subject(s) - human leukocyte antigen , biology , single nucleotide polymorphism , allele , snp , microrna , population , genetics , antigen , microbiology and biotechnology , immunology , genotype , gene , medicine , environmental health
Surface expression of human leukocyte antigen (HLA)‐class I molecules is critical for modulating T/natural killer lymphocytes' effector functions. Among HLA molecules, HLA‐C, the most recently evolved form of class I antigens, is subjected to both transcriptional and multiple post‐transcriptional regulation mechanisms affecting its cell surface expression. Among the latter a region placed in the 3′ untranslated region of HLA‐C transcript contains the single nucleotide polymorphism (SNP) rs67384697 “G‐ins/del” that has been found to be strictly associated with surface levels of HLA‐C allomorphs because of the effect on the binding site of a microRNA (Hsa‐miR‐148a). Higher expression of HLA‐C has been proved to influence HIV‐1 infection via a better control of viremia and a slower disease progression. More importantly, the analysis of SNP rs67384697 “G‐ins/del” combined with the evaluation of the HLA‐Bw4/‐Bw6 C1/C2 supratype, as well as the killer immunoglobulin‐like receptor genetic asset, has proved to be pivotal in defining the status of Elite Controllers in the Caucasian population. Here we describe a new reliable and fast method of allele‐specific real‐time PCR to monitor the integrity/disruption of the binding site of the microRNA Hsa‐miR‐148a in a high‐throughput format that can be easily applied to studies involving large cohorts of individuals.