Project description:AGO-iCLIP-seq was utilized to characterize the RNAs bound to AGO in human neural stem cells infected with Zika virus strains Paraiba at MOI 1. miRNAs and mRNAs were analyzed to determine changes in RNAs loaded into the RISC 4 days post-infection.
Project description:Viral interfering RNA (viRNA) has been identified from several viral genomes via directly deep RNA sequencing of the virus-infected cells, including zika virus (ZIKV). Once produced by endoribonuclease Dicer, viRNAs, similar to microRNAs, are loaded onto Argonaute (AGO) family proteins of the RNA-induced silencing complexes (RISCs) to pair with their RNA targets and then initiate cleavage of the target genes. However, identities of functional ZIKV viRNAs and their viral RNA targets remain largely unknown. By combining AGO-associated RNA sequencing, deep sequencing analysis in ZIKV-infected neural stem cells (NSCs), and miRanda target scanning, we have defined 29 ZIKV derived viRNA profiles in NSCs, and established the complex interaction networks between the viRNAs and their viral targets. Our recent study has shown that ZIKV capsid protein interacted with Dicer and antagonized its endoribonuclease activity depending on its histidine (H) at the 41st amino acid. Accordingly, the rescued ZIKV-H41R mutant virus, compared to wild-type ZIKV, no longer suppressed Dicer enzymatic activity and consequently failed to inhibit miRNA biogenesis in NSCs. As a result, much higher levels of viRNAs generated from the ZIKV-H41R virus-infected NSCs, suggesting Dicer-dependent viRNA production. Knockdown of individual RNAi machinery in ZIKV-infected NSCs suggests that viRNA is a limiting factor of ZIKV infection in NSCs. The mapping of viRNAs to their RNA targets is paving a way to further investigate how viRNAs play the role in anti-viral mechanisms or even other unknown biological functions.
Project description:RNA-seq was utilized to characterize the transcriptome of human neural stem cells infected with Zika virus strains MR766 and Paraiba at MOI 1. Coding and noncoding genes were analyzed to determine transcriptional changes 3 days post-infection.
Project description:small RNA-seq was utilized to characterize the transcriptome of human neural stem cells infected with Zika virus strains MR766 and Paraiba at MOI 1. Coding and noncoding genes were analyzed to determine transcriptional changes 3 days post-infection.
Project description:Nine patients with temporal lobe epilepsy (TLE) underwent hippocampal resection. Six had hippocampal sclerosis, three did not show signs of hippocampal sclerosis. Resected hippocampi were used for Ago iCLIP to detect miRNA targeting in the resected tissue.
Project description:Purpose: The goal of this study is to investigate the effect of Zika virus infection on neural stem cells Methods: RNA-Seq transcriptome analysis of neural stem cells infected with Zika virus compared to mock infected