https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/329 Sat, 04 Jul 2026 09:10:54 GMT 2026-07-04T09:10:54Z https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/210903 Title: Oxytocin Modulates Microglial IL-17-Linked Inflammatory Pathways Through the IL-6/COX-2 Authors: Hwang, Woochang; Jang, Yong-hun; Hong, Juyoung; Kang, Suyeon; Hur, Junho K.; Lee, Hyunju Abstract: Neonatal neuroinflammation, driven by microglial activation and cytokine signaling, contributes to brain injury and adverse neurodevelopment outcomes. Perinatal inflammatory mediators, including interleukin-6, cyclooxygenase-2, and interleukin-17, prime microglia and influence circuit vulnerability. This study investigated whether oxytocin pretreatment attenuates lipopolysaccharide-induced inflammatory priming in BV-2 microglial cells. BV-2 microglia were preincubated with oxytocin (33 ng/mL) for 2 h, followed by lipopolysaccharide (0.5 µg/mL) for 2 h. Expression of ionized calcium-binding adapter molecule 1, a microglia marker, in BV-2 cells was assessed by immunofluorescence. After lipopolysaccharide treatment, the gene expression of BV-2 cells was assayed at 1, 2, and 6 h post stimulation by RT-qPCR and RNA-seq. Functional characterization of gene expression profile was performed. Analyses of gene expression profile of BV-2 cells by RT-qPCR and RNA-seq revealed that oxytocin pretreatment attenuated lipopolysaccharide-induced transcriptional activation, including interleukin-6 and cyclooxygenase-2 upregulation. Pathway enrichment analyses suggested that oxytocin-responsive genes were linked to the interleukin-17 signaling pathway. Gene Ontology enrichment analysis showed enrichment for genes related to cytokine production, membrane raft, and chemokine activity. Oxytocin pretreatment mitigates lipopolysaccharide-induced microglial activation by modulating the interleukin-17–interleukin-6/cyclooxygenase-2 axis, suggesting its potential role for oxytocin as an endogenous modulator of neuroinflammation during early brain development. Thu, 01 Jan 2026 00:00:00 GMT https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/210903 2026-01-01T00:00:00Z https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/209322 Title: Ultrasensitive single-point mutation detection via digital counting of individual dCas9-DNA complexes Authors: Kim, Seohyun; Lee, Wi-jae; Lee, Hyomin K.; Ji, Sangmin; Hur, Junho K.; Lee, Seung Hwan; Koh, Hye-ran Abstract: Single-point DNA mutations represent the majority of oncogenic mutations and contribute significantly to genetic variations. Thus, ultrasensitive methods to detect a very low fraction of single-point mutations are critical for the early diagnosis of mutation-associated genetic diseases. Here, we developed a highly sensitive approach for detecting single-point mutations by counting individual nuclease-dead Cas9 (dCas9) complexes bound to DNA at the single-molecule level. The clustered regularly interspaced short palindromic repeats (CRISPR)/dCas9 system, composed of dCas9 protein and guide RNA (gRNA), specifically recognizes DNA sequences complementary to the gRNA and bearing an appropriate protospacer adjacent motif (PAM) adjacent to the target site. By combining the PAM-dependent binding specificity with single-molecule counting, we successfully discriminated single-point mutations in clinically relevant genes, including EGFR (c.2573 T > G) and KRAS (c.34 G>T and c.35 G>A). Kinetic and thermodynamic analyses revealed that the discrimination mechanism was primarily driven by the faster association rate of the dCas9-gRNA complex to target DNA with a canonical PAM. Furthermore, we quantitatively measured the mutant allele fraction across the entire range of mutation rates and achieved discrimination of mutations present at levels as low as 0.5 %. These results suggest that digital single-molecule counting of dCas9 complexes may serve as a promising approach for early and sensitive detection of single-point mutations. Mon, 01 Dec 2025 00:00:00 GMT https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/209322 2025-12-01T00:00:00Z https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/208342 Title: SARS-CoV-2 RNA-binding protein suppresses extracellular miRNA release Authors: Mun, Hyejin; Shin, Chang Hoon; Fei, Qingxuan; Giraldo, Andrea Estefania Lopez; Choi, Kyoung-Min; Lee, Ji Won; Kim, Kyungmin; Min, Kyung-Won; Shi, Leilei; Bedford, Mark T.; Kim, Dong-Chan; Chun, Yoo Lim; Ryu, Seonghyun; Kim, Dongin; Chang, Jeong Ho; Westrope, Ryan T.; Shay, Michelle; Nguyen, Edward; Hur, Junho K.; Agyenda, Abigail; Kim, Nam Chul; Kang, Sung-Ung; Lee, Woonghee; Yoon, Je-Hyun Abstract: SARS-CoV-2 is the betacoronavirus causing the COVID-19 pandemic. Although the SARS-CoV-2 genome and transcriptome were reported previously, the function of individual viral proteins is largely unknown. Utilizing biochemical and molecular biology methods, we identified that four SARS-CoV-2 RNA-binding proteins (RBPs) regulate the host RNA metabolism by direct interaction with mature miRNA let-7b revealed by Nuclear Magnetic Resonance spectroscopy (NMR). SARS-CoV-2 RBP Nsp9 primarily binds mature miRNA let-7b, a direct ligand of the Toll-like Receptor 7 (TLR7), one of the potential SARS-CoV-2 therapeutics. Nsp9 suppresses host gene expression possibly by promoting let-7b-mediated silencing of a cellular RNA polymerase, POLR2D. In addition, Nsp9 inhibits extracellular release of let-7b and subsequent antiviral activity via TLR7. These results demonstrate that SARS-CoV-2 hijacks the host RNA metabolism to suppress antiviral responses and to shut down cellular transcription. Our findings of how a natural ligand of TLR7, miRNA let-7b, is suppressed by SARS-CoV-2 RBPs will advance our understanding of COVID-19 and SARS-CoV-2 therapeutics. Mon, 01 Dec 2025 00:00:00 GMT https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/208342 2025-12-01T00:00:00Z https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/212123 Title: shRNAI: A deep neural network for the design of highly potent shRNAs Authors: Park, Seokju; Park, Seong-Ho; Oh, Jin-Seon; Hong, Sumin; Been, Kyung Wook; Noh, Yung-Kyun; Hur, Junho K.; Nam, Jin-Wu Abstract: miRNA-mimicking short hairpin RNAs (shRNAmirs), which depend on the endogenous miRNA biogenesis pathway, have been widely used to investigate gene function and to develop therapeutic strategies due to their stable and robust knockdown of target genes. However, despite the efforts to design potent shRNAmir guide RNAs (gRNAs), relevant biological features beyond the primary sequence have not been fully explored. Here, we present shRNAI, a convolutional neural network model for predicting highly potent shRNAmir gRNAs. Even when trained solely on gRNA sequences, shRNAI outperforms previous algorithms. We further improved the model (shRNAI+) by adding features related to shRNAmir processability and target site context, resulting in superior performance across both public datasets and our own experimental tests. Although shRNAI was initially trained on datasets built with a CNNC motif-free pri-miR-30 backbone, it also displayed improved performance on the CNNC motif. Overall, our study provides a robust framework for designing potent shRNAmir gRNAs, as well as a versatile tool for developing RNAi therapeutics. Mon, 01 Dec 2025 00:00:00 GMT https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/212123 2025-12-01T00:00:00Z