Evaluating NDUFA4's Role in Zika Virus Replication
ZIKV replicons have been previously reported (Rusanov et al., 2018 (link)). To determine whether NDUFA4 is involved in flavivirus genome replication, WT, NDUFA4−/− and NDUFA4ΔSNP iPSCs were transfected with the in vitro transcribed ZIKV replicon RNA. iPSCs were plated into 24-well plates at 62,500 cells per well. Transfection mixes for each well were prepared by mixing 0.1 μg of the replicon RNA with 0.2 μL each of the mRNA boost reagent and TransIT-mRNA reagent (TransIT-mRNA transfection kit; Mirus, WI). At 24 hpi, cells were lysed in 1X cell culture lysis reagent (Promega, WI) followed by measurement of the Renilla luciferase activity.
Han Y., Tan L., Zhou T., Yang L., Carrau L., Lacko L.A., Saeed M., Zhu J., Zhao Z., Nilsson-Payant B.E., Lira Neto F.T., Cahir C., Giani A.M., Chai J.C., Li Y., Dong X., Moroziewicz D., Paull D., Zhang T., Koo S., Tan C., Danziger R., Ba Q., Feng L., Chen Z., Zhong A., Wise G.J., Xiang J.Z., Wang H., Schwartz R.E., tenOever B.R., Noggle S.A., Rice C.M., Qi Q., Evans T, & Chen S. (2022). A human iPSC-array-based GWAS identifies a virus susceptibility locus in the NDUFA4 gene and functional variants. Cell Stem Cell, 29(10), 1475-1490.e6.
Other organizations :
Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Memorial Sloan Kettering Cancer Center, New York University, Rockefeller University, Boston University, Instituto de Medicina Integral Professor Fernando Figueira, Tri-Institutional PhD Program in Chemical Biology, Albert Einstein College of Medicine, New York Stem Cell Foundation, Shanghai Jiao Tong University, Central China Normal University
Renilla luciferase activity (as a measure of ZIKV replicon RNA replication)
control variables
Amount of replicon RNA transfected per well (0.1 μg)
Amount of mRNA boost reagent and TransIT-mRNA reagent per well (0.2 μL each)
Cell seeding density (62,500 cells per well)
Time point of cell lysis (24 hpi)
controls
Positive control: Not specified
Negative control: Not specified
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to
get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
