Epitranscriptomic N-6-Methyladenosine Modification Is Required for Direct Lineage Reprogramming into Neurons
ACS CHEMICAL BIOLOGY
Authors: Choi, Hwan; Baek, Soonbong; Cho, Byounggook; Kim, Siyoung; Kim, Junyeop; Chang, Yujung; Shin, Jaein; Kim, Jongpil
Abstract
N-6-methyladenosine (m(6)A), a conserved epitranscriptomic modification of eukaryotic mRNA (mRNA), plays a critical role in a variety of biological processes. Here, we report that m(6)A modification plays a key role in governing direct lineage reprogramming into induced neuronal cells (iNs). We found that m(6)A modification is required for the remodeling of specific mRNAs required for the neuronal direct conversion. Inhibition of m(6)A methylation by Mettl3 knockdown decreased the efficiency of direct lineage reprogramming, whereas increased m(6)A methylation by Mettl3 overexpression increased the efficiency of iN generation. Moreover, we found that transcription factor Btg2 is a functional target of m(6)A modification for efficient iN generation. Taken together, our results suggest the importance of establishing epitranscriptomic remodeling for the cell fate conversion into iNs.
A fast and reliable method for detecting SNP rs67384697 (Hsa-miR-148a binding site) by a single run of allele-specific real-time PCR
HLA
Authors: Malnati, Mauro S.; Biswas, Priscilla; Ugolotti, Elisabetta; Di Marco, Eddi; Sironi, Francesca; Parolini, Francesca; Garbarino, Lucia; Mazzocco, Michela; Zipeto, Donato; Biassoni, Roberto
Abstract
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 30 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.