TY  - JOUR
TI  - Quantification of Extracellular Double-stranded RNA Uptake and Subcellular Localization  Using Flow Cytometry and Confocal Microscopy
AU  - Nguyen, Tan A
AU  - Whitehead, Lachlan 
AU  - Pang, Ken C. 
VL  - 8
IS  - 12
PY  - 2018
DA  - 2018/06/20
SP  - e2890
C1  - Bio-protocol 2018;8:e2890
DO  - 10.21769/BioProtoc.2890
UR  - https://doi.org/10.21769/BioProtoc.2890
AB  - Double-stranded RNA is a potent pathogen-associated molecular pattern (PAMP) produced as a by-product of viral replication and a well-known hallmark of viral infection. Viral dsRNAs can be released from infected cells into the extracellular space and internalized by neighboring cells via endocytosis. Mammals possess multiple pattern recognition receptors (PRRs) capable of detecting viral dsRNAs such as endosomal toll-like receptor 3 (TLR3) and cytosolic RIG-I-like receptors (RLRs) which lead to the production of type I interferons (IFNs). Thus, intracellular localization of viral dsRNA can provide insight into the downstream signaling pathways leading to innate immune activation. Here, we describe a quantitative method for measuring extracellular dsRNA uptake and visualizing subcellular localization of internalized dsRNA via flow cytometry and confocal microscopy respectively.
KW  - Double-stranded RNA
KW  - Endosomes
KW  - Lysosomes
KW  - Viruses
KW  - Poly(I:C)
KW  - TLR3
KW  - RIG-I
KW  - MDA-5
KW  - Confocal microscopy
KW  - Flow cytometry
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False