Zika virus (ZIKV) is an emergent member of the Flaviviridae family which causes severe congenital defects and other major sequelae, but the cellular processes that support ZIKV replication are incompletely understood. Related flaviviruses use the endoplasmic reticulum (ER) as a membranous platform for viral replication and induce ER stress during infection. Our data suggest that ZIKV activates IRE1α, a component of the cellular response to ER stress. IRE1α is an ER-resident transmembrane protein that possesses a cytosolic RNase domain. Upon activation, IRE1α initiates nonconventional cytoplasmic splicing of XBP1 mRNA. Spliced XBP1 encodes a transcription factor, which upregulates ER-related targets. We find that ZIKV infection induces XBP1 mRNA splicing and induction of XBP1 target genes. Small molecule inhibitors of IRE1α, including those specific for the nuclease function, prevent ZIKV-induced cytotoxicity, as does genetic disruption of IRE1α. Optimal ZIKV RNA replication requires both IRE1α and XBP1. Spliced XBP1 has been described to cause ER expansion and remodeling and we find that ER redistribution during ZIKV infection requires IRE1α nuclease activity. Finally, we demonstrate that inducible genetic disruption of IRE1α and XBP1 impairs ZIKV replication in a mouse model of infection. Together, our data indicate that the ER stress response component IRE1α promotes ZIKV infection via XBP1 and may represent a potential therapeutic target.proteins: protein kinase receptor-like ER kinase (PERK), activating transcription factor 6 (ATF6), and inositol-requiring enzyme 1 α (IRE1α). ER stress causes IRE1α to undergo oligomerization and autophosphorylation, which activates its cytosolic RNase domain to initiate nonconventional splicing of XBP1 mRNA. Spliced XBP1 is a specific product of activated IRE1α and encodes a transcription factor that upregulates targets that are involved in ER function [11]. IRE1α also targets other specific RNAs, leading to their degradation in a process termed regulated IRE1-dependent decay (RIDD) [12,13].The role of IRE1α in infection appears to vary for different members of the Flaviviridae family. Hepatitis C virus (HCV) activates IRE1α [14] to promote viral replication [15] independently of XBP1 by preventing apoptotic death of infected cells [16]. Dengue (DENV) and Japanese encephalitis viruses (JEV) also benefit from IRE1α via an XBP1-independent mechanism [17-20], whereas West Nile virus (WNV) replication is unaffected by either IRE1α [21,22] or XBP1 [23]. Conflicting results have been obtained for tick-borne encephalitis virus, with IRE1α nuclease inhibition either limiting viral replication [24] or having no effect [22]. ZIKV activates IRE1α, as demonstrated by the presence of spliced XBP1 in ZIKV-infected cultured cells and brain tissue from ZIKV-infected embryonic mice [25][26][27]. In this study, we examined the role of IRE1α in ZIKV infection and found that IRE1α promotes ZIKV replication via XBP1 in cultured cells. We further found that genetic disruption of IRE1α and XBP1 lim...

Preclass reading quizzes (RQs) have been shown to enhance student performance. Many instructors implementing evidence-based teaching assign preclass RQs to ensure their students are prepared to engage in class activities. Textbook companies now offer a gamified, adaptive-learning RQ format. In these RQs, students answer point-valued questions until they reach a threshold. If students answer incorrectly, the question decreases in point value on the next attempt. These RQs also give students who answer questions incorrectly more questions on that topic and direct students to sections of a textbook they need to review. We assessed the impact of gamified, adaptive preclass RQs compared with more traditional preclass RQs on in-class RQs and course exam performance as well as students’ perceptions of RQs. Students in the gamified, adaptive treatment performed equally compared with students in the traditional, static treatment on in-class RQs and course exams. While students in the gamified, adaptive treatment did have a more positive perception of preclass RQs, this factor explained less than 3% of the variation in RQ perception. Our findings suggest that instructors should verify that gamified, adaptive technologies impact student learning in their course before integrating them into their course and asking students to pay for them.