The NSs protein is a major virulence factor in bunyaviruses, crucial for viral pathogenesis. However, assessing NSs protein function can be challenging due to its inhibition of cellular RNA polymerase II, impacting NSs protein expression from plasmid DNA. The recombinant Rift Valley fever virus (RVFV) MP-12 strain (rMP-12), a highly attenuated vaccine strain, can be safely manipulated under biosafety level 2 conditions. Leveraging a reverse genetics system, we can engineer rMP-12 variants expressing heterologous NSs genes, enabling functional testing in cultured cells. Human macrophages hold a central role in viral pathogenesis, making them an ideal model for assessing NSs protein functions. Consequently, we can comprehensively compare and analyze the functional significance of various NSs proteins in human macrophages using rMP-12 NSs variants. In this chapter, we provide a detailed overview of the preparation process for rMP-12 NSs variants and introduce two distinct human macrophage models: THP-1 cells and primary macrophages. This research framework promises valuable insights into the virulence mechanisms of RVFV and other bunyaviruses and the potential for vaccine development.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1007/978-1-0716-3926-9_25 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Periodic Light Modulations for Low-Cost Wide-Field Imaging of Luminescence Kinetics Under Ambient Light.
Adv Sci (Weinh)
January 2025
PASTEUR, Département de chimie, École normale supérieure, PSL University, Sorbonne Université, CNRS, 24, rue Lhomond, Paris, 75005, France.
Imaging luminescence kinetics is invaluable in many fields, including biology and chemistry. However, the luminescence lifetime of most photo-activated states is in the low ns-µs range and its measurement requires adding costly image intensifiers to cameras to access the fast phenomena present. Here, the Rectified Imaging under Optical Modulation (RIOM) and Heterodyne Imaging under Optical Modulation (HIOM) protocols make this possible with standard low-cost cameras only, even under ambient light.
View Article and Find Full Text PDFInvestigating the reassortment potential and pathogenicity of the S segment in Akabane virus using a reverse genetics system.
BMC Vet Res
January 2025
Laboratory of Veterinary Infectious Disease, College of Veterinary of Medicine, Jeonbuk National University, Iksan, Jeonbuk, 54596, Republic of Korea.
Background: Akabane virus (AKAV) is an arthropod-borne virus that causes congenital malformations and neuropathology in cattle and sheep. In South Korea, AKAVs are classified into two main genogroups: K0505 and AKAV-7 strains. The K0505 strain infects pregnant cattle, leading to fetal abnormalities, while the AKAV-7 strain induces encephalomyelitis in post-natal cattle.
View Article and Find Full Text PDFProtein Fusion of Biosynthetic Enzymes and a Thermo-Responsive Polypeptide Expedites Facile Access to Biocatalysts for Nucleotide Sugars.
Chembiochem
January 2025
Shandong University - Qingdao Campus, National Glycoengineering Research Center, Room 230, Ganchang Yard F Block, Qingdao campus of Shandong University, 72 Binhai Road,, Jimo District, Qingdao, Shandong, 266237 China, 266237, Qingdao, CHINA.
Nucleotide sugars (NSs) are essential building blocks for the enzymatic assembly of glycans. In this study, we established a preparation and recycling avenue to the biocatalysts for the enzymatic synthesis of NSs. This approach involves fusing two enzymes into a bifunctional chimera and using elastin-like polypeptides (ET64) as a purification tag, which allows for easy recovery of these biocatalysts without the need for chromatography.
View Article and Find Full Text PDFRBBP6 anchors pre-mRNA 3' end processing to nuclear speckles for efficient gene expression.
Mol Cell
January 2025
Department of Microbiology and Molecular Genetics, School of Medicine, University of California, Irvine, Irvine, CA 92697, USA. Electronic address:
Pre-mRNA 3' processing is an integral step in mRNA biogenesis. However, where this process occurs in the nucleus remains unknown. Here, we demonstrate that nuclear speckles (NSs), membraneless organelles enriched with splicing factors, are major sites for pre-mRNA 3' processing in human cells.
View Article and Find Full Text PDFAstrocytic NLRP3 cKO mitigates depression-like behaviors induced by mild TBI in mice.
Neurobiol Dis
January 2025
Department of Anesthesiology, Hebei Province, Cangzhou Hospital of Integrated Traditional and Western Medicine, Cangzhou, China,. Electronic address:
Background: Reports indicate that depression is a common mental health issue following traumatic brain injury (TBI). Our prior research suggests that Nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3)-related neuroinflammation, modulated by glial cells such as astrocytes, is likely to play a crucial role in the progression of anxiety and cognitive dysfunction. However, there is limited understanding of the potential of astrocytic NLRP3 in treating depression under mild TBI condition.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!