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Amplification of immunity by engineering chicken MDA5 combined with the C terminal domain (CTD) of RIG-I
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Journal Article
Amplification of immunity by engineering chicken MDA5 combined with the C terminal domain (CTD) of RIG-I
2022
Overview
Innate immune system is triggered by pattern recognition receptors (PRRs) recognition. Retinoic acid-inducible gene 1 (RIG-I) is a major sensor that recognizes RNA ligands. However,
chickens
have no homologue of RIG-I; instead, they rely on melanoma differentiation-associated protein 5 (MDA5) to recognize RNA ligands, which renders
chickens
susceptible to infection by influenza A viruses (IAVs). Here, we engineered the
cMDA5
viral RNA sensing domain (C-terminal domain, CTD) such that it functions similarly to
human RIG-I
(
hRIG-I
) by mutating histidine 925 into phenylalanine, a key residue for
h
RIG-I RNA binding loop function, or by swapping the CTD of
cMDA5
with that of
hRIG-I
or
duck RIG-I
(
dRIG-I
). The engineered
cMDA5
gene was expressed in
cMDA5
knockout DF-1 cells, and interferon-beta (
IFN-β
) activity and expression of interferon-related genes were measured after transfection of cells with RNA ligands of
h
RIG-I or
human
MDA5 (
h
MDA5). We found that both mutant
cMDA5
and engineered
cMDA5
triggered significantly stronger interferon-mediated immune responses than wild-type
cMDA5
. Moreover, engineered
cMDA5
reduced the IAV titer by 100-fold compared with that in control cells. Collectively, engineered
cMDA5/RIG-I CTD
significantly enhanced interferon-mediated immune responses, making them invaluable strategies for production of IAV-resistant
chickens
.
Key points
• Mutant chicken MDA5 with critical residue of RIG-I (phenylalanine) enhanced immunity.
• Engineered chicken MDA5 with CTD of RIG-I increased IFN-mediated immune responses.
• Engineered chicken MDA5 reduced influenza A virus titers by up to 100-fold.
Publisher
Springer Berlin Heidelberg,Springer,Springer Nature B.V
Subject
/ Animals
/ Applied Microbial and Cell Physiology
/ Biomedical and Life Sciences
/ Chickens
/ DEAD-box RNA Helicases - genetics
/ DEAD-box RNA Helicases - metabolism
/ Domains
/ Ducks
/ genes
/ Homology
/ Humans
/ Immunity
/ Interferon-Induced Helicase, IFIH1 - genetics
/ Ligands
/ Melanoma
/ Methods
/ Microbial Genetics and Genomics
/ Mutants
/ Pattern recognition receptors
/ Poultry
/ Residues
/ RNA
