SLC35A1 Pre-design Chimera RNAi
- Known as:
- SLC35A1 Pre-design Chimera RNAi
- Catalog number:
- H00010559-R01
- Product Quantity:
- 10 nmol
- Category:
- -
- Supplier:
- Abno
- Gene target:
- SLC35A1 Pre-design Chimera RNAi
Related genes to: SLC35A1 Pre-design Chimera RNAi
- Gene:
- SLC35A1 NIH gene
- Name:
- solute carrier family 35 member A1
- Previous symbol:
- -
- Synonyms:
- CMPST, hCST
- Chromosome:
- 6q15
- Locus Type:
- gene with protein product
- Date approved:
- 1999-10-05
- Date modifiied:
- 2016-02-17
Related products to: SLC35A1 Pre-design Chimera RNAi
Related articles to: SLC35A1 Pre-design Chimera RNAi
- Avian influenza virus (AIV) poses a persistent threat to poultry health and food security, with conventional control measures offering limited protection. A promising alternative is the use of gene editing to generate host resistance by ablating viral entry receptors or cellular proteins that are required for completion of the viral life cycle. The solute carrier family 35 member A1 (SLC35A1) gene encodes a Golgi-localized CMP-sialic acid transporter that is a key step in the sialylation of glycoproteins. In this study, we used the CRISPR/Cas9 system to disrupt SLC35A1 in chicken DF-1 fibroblasts and evaluated the effect on sialic acid expression and susceptibility to different strains of AIV. Lectin staining and flow cytometry confirmed a significant reduction in α2,3-linked sialic acids in SLC35A1 knockout cells, while α2,6-linked sialic acids were absent in the cells regardless of genotype. Infection experiments with three avian influenza virus strains (H5N1/PR8, H5N2, and H7N1) revealed that SLC35A1 knockout reduced viral replication in a strain-specific manner. Knockout cells infected with H5N1/PR8 showed the greatest dependence on SLC35A1-mediated sialylation with decreased viral load at 24 hours post-infection (hpi) and 48 hpi compared to wildtype cells and no observable viral growth between the timepoints. Infection of knockout cells with H5N2 resulted in a modest decrease in viral load at both timepoints as well as absence of viral growth. On the other hand, infection of knockout cells with H7N1 resulted in decreased viral load only at 48 hpi compared to wildtype cells, but the amount of virus in knockout cultures increased from 24 hpi to 48 hpi. These results demonstrate that SLC35A1 is a key host factor that supports AIV entry via α2,3-linked sialic acids; however, viral dependency on this host factor may be confounded by strain. - Source: PubMed
Publication date: 2026/05/25
Liu HeidiCruvinel Jéssica MoraesWarren Wesley CMa WenjunChen Paula R - Controlling the generation of toxic by-products in mammalian bioprocess to maximize therapeutic protein production and glycosylation patterns is a challenge. Intracellular metabolism is often not well-regulated and known to secrete toxic intermediate by-products which hampers cellular performance and negatively impacts critical quality attributes (CQA) of cells. Previous studies have identified trigonelline (TRI), n-acetyl putrescine (NAP), aconitic acid (AA), and cytidine monophosphate (CMP) generated through CHO cell metabolism and verified their negative impacts on growth and antibody production. In this approach, a genetic engineering strategy was developed to control downstream accumulation of inhibitory metabolites. The study successfully identified four different metabolic genes in CHO cells, including Cat (nicotinate and nicotinamide metabolism) to control the generation of TRI, Got1 and Hoga1 (proline metabolism) to control the generation of NAP, Got1 (TCA cycle) to control the generation of AA, and Slc35a1 (n-glycan biosynthesis) to control the generation of CMP. Each target gene-of-interest (GOI) was cloned from CHO genomic library, inserted into linearized vector plasmid, and subsequently transfected into cells. CQA of the bioprocess realized 22-30% increase in peak cell density, 16-22% increase overall IVCD, with an improving growth rate during cellular expansion phase when comparing engineered cells against control cells. The study also conducted a follow-up quadruple transfection study where all four GOIs were co-transfected into cells at ¼ of the total DNA concentration per GOI. An increase in cellular performance was also realized, as increases in peak VCD (17% increase), cumulative IVCD (17% increase), and growth rate were achieved. Both studies also found higher IgG1 antibody synthesis when cell metabolism was better regulated, as the studies measured 4% to 40% titer increase across all engineered cells when compared against control cells. The study also measured higher levels of G1F and G2F glycans with decreased level of G0F across all transfected cells, further indicating improvement in bioprocess, as cells were able to produce a higher fraction of semi-complex and complex versus simple glycoforms. Further investigation revealed that Cat and Slc35a1 exhibited comparable expression levels in the MG condition to their single-gene conditions (within 1% and 10% difference, respectively), corresponding to modest titer improvements closest to the control. These findings suggest that when all four genes are co-expressed, Cat and Got1 may act as rate-limiting factors influencing both cellular phenotypes and titer production. In both studies, the concentrations of downstream metabolic inhibitors were measured to be significantly decreased when comparing engineered cells against control cells, further demonstrating that overexpression of genes to re-allocate metabolic fluxes away from synthesizing toxic by-products can significantly improve cellular growth and protein synthesis. - Source: PubMed
Publication date: 2026/05/20
Hoang DucKuang BingyuWang ZhaoLiang GeorgeYoon Seongkyu - Bovine parainfluenza virus type 3 (BPIV3) is a leading cause of respiratory illness in cattle and a primary component of the bovine respiratory disease complex (BRDC), resulting in significant economic losses. Understanding the mechanisms of BPIV3 infection, particularly the entry process, is essential for developing effective control measures. Identifying specific host factors that viruses exploit during their life cycle can reveal critical vulnerabilities for potential antiviral targets. We established a genome-wide CRISPR/Cas9 knockout screen in bovine cells to identify host factors involved in viral infections. Our screen identified several key host factors required for BPIV3 infection, including the sialic acid transporter SLC35A1 and the Sm-like protein LSM12. Further mechanistic analysis revealed that these factors played critical roles at distinct stages of the BPIV3 entry process. These findings not only advance our understanding of how BPIV3 infects host cells but also identify potential host targets for inhibiting infection and developing novel antiviral strategies. - Source: PubMed
Publication date: 2026/01/28
Hao JinyangGao XiaoranLight ChristineSun YuzeLu ShaTian YuGao XiaSu YuanGao JieHuang XinZhang QianyiWang JinliangHai RongHu WeiWang Guojun - BACKGROUND: The ependymal glycocalyx (Gcx) is a glycan-rich apical structure that lines the ventricular brain surface. It is thought to contribute to cerebrospinal fluid dynamics and brain homeostasis by forming a selective barrier, preserving surface charge, and supporting ciliary function. Despite its importance, the structural integrity and glycan composition of the ependymal Gcx remain poorly understood, particularly in the context of physiological aging and acute neurological injury, such as intraventricular hemorrhage (IVH). We aimed to elucidate the physiological role of the ependymal Gcx and its alterations in response to aging and acute brain injury. METHODS: We comprehensively investigated age- and injury-related changes in the ependymal Gcx using young (8–10-week-old), aged (60–62-week-old), and IVH model mice. The Gcx structure was visualized using lanthanum-enhanced electron microscopy, and glycan profiles were assessed through double immunofluorescence staining with S100β and a panel of 21 fluorescent lectins. Gcx thickness was quantitatively analyzed using a novel image analysis approach based on fluorescence intensity profiles. Single-cell RNA sequencing (scRNA-seq) was performed on ventricular tissue to identify transcriptional changes in aged ependymal cells related to glycan biosynthesis, glycan sialylation, desialylation, vesicular transport, and inflammatory responses. RESULTS: Immunohistochemistry showed that, in young mice, the ependymal Gcx was bound to SiaFind and Lycopersicon esculentum lectin; additionally, it is bound to PNA even without desialylation. In aged mice, the Gcx displayed marked thinning, detachment, and significant loss of terminal sialic acids. In young mice, Gcx disruption after IVH peaked on day 3 and correlated with periventricular inflammation; in contrast, the inflammation persisted in aged IVH mice. Integrated single-cell RNA-seq revealed age-related alterations. Key sialylation genes (ST3GAL1, SLC35A1) and core 1 O-glycan enzymes (C1GALT1, C1GALT1C1) were downregulated, whereas ST3GAL5 and plasma-membrane sialidase NEU3 were upregulated. Additionally, senescence markers (Cdkn1a, Trp53) and multiple interferon-stimulated genes were elevated. CONCLUSIONS: The ependymal Gcx is a dynamic and injury-sensitive structure whose integrity is compromised by aging and IVH. Its disruption promotes neuroinflammation and may contribute to the development of hydrocephalus and neurodegeneration. Therapeutic modulation of glycosylation pathways may provide a promising strategy to preserve Gcx function and protect the internal brain environment. - Source: PubMed
Publication date: 2025/11/07
Iida TomohiroMori KosukeTomita HiroyukiOhmura KazufumiTaguchi KohtaroNiwa AyumiKanayama TomohiroSugie ShigeyukiOkada HideshiIzumo TsuyoshiHara Akira - Porcine epidemic diarrhea virus (PEDV) is a globally distributed alphacoronavirus with economic importance that can cause severe watery diarrhea and even death in piglets. To identify host factors essential for PEDV infection, we performed a genome-wide CRISPR/Cas9 screen in human hepatocellular carcinoma cells (Huh7) using the highly virulent PEDV GIIb strain GDU. Several genes involved in the sialic acid and heparan sulfate biosynthesis pathway and cholesterol metabolism were highly enriched following PEDV selection. We validated that the host factor ST3 beta-galactoside alpha-2,3-sialyltransferase 4 (ST3GAL4), which catalyzes the transfer of sialic acid to sugar chains via α2,3-linked linkages, is important for PEDV infection. To systematically investigate the role of sialic acid in PEDV infection, we knocked out genes related to sialic acid synthesis. This led to a reduced abundance of sialic acid on the cell surface, which in turn inhibited PEDV adsorption and internalization. Furthermore, we found that both α2,3-linked and α2,6-linked sialic acids can serve as cellular attachment factors for PEDV. We conducted a glycan microarray screen to determine which sialoglycans are preferred by the PEDV spike protein. The results revealed that PEDV favors binding to α2,3-sialoglycans. Additionally, we found that not only current circulating PEDV strains but also other porcine coronaviruses rely on sialic acid for efficient infection. Collectively, our findings provide insights into critical host factors involved in PEDV infection and demonstrate that disrupting genes involved in sialic acid biosynthesis negatively affects the infectivity of multiple porcine enteric coronaviruses.IMPORTANCEA wide range of viruses utilize sialic acid as receptors. Sialic acid binding may serve as a key determinant of viral host range. Different viruses exhibit distinct preferences for specific types of sialic acid linkages. However, it remains unclear which specific subtypes of sialic acid are utilized during PEDV infection. In this study, we performed CRISPR-based genome-wide knockout screening and identified ST3GAL4 as a key host factor for PEDV infection. Furthermore, we found that both α2,3-linked and α2,6-linked sialic acids can function as attachment factors for PEDV infection. A glycan microarray screen revealed that PEDV S1 shows the strongest binding preference for α2,3-linked and α2,8-linked sialosides. Sialic acids were also implicated in infections by other porcine enteric coronaviruses. Overall, our findings advance our understanding of viral entry mechanisms of PEDV and other swine coronaviruses and may provide avenues for designing antiviral strategies. - Source: PubMed
Publication date: 2025/08/06
Guo GuanghaoZhang MengjiaXu ZhuojiaXi PengZhu HongmeiEvers AnoukLebbink Robert JanLang YifeiHe QigaiHuang Yao-WeiLi TiehaiBosch Berend JanLi Wentao