TY - JOUR
T1 - Shedding light on microRNA function via microscopy-based screening
AU - Rodrigues Lopes, Ines
AU - Silva, Ricardo Jorge
AU - Caramelo, Ines
AU - Eulalio, Ana
AU - Mano, Miguel
N1 - Funding Information:
The authors thank L. Braga (ICGEB, Italy) for assistance with the widefield/confocal image acquisition and R. Hollandi and P. Horvath (BRC, Hungary) for critical input on the CellProfiler workflow. R.J.S. is a recipient of a PhD fellowship of the Doctoral Programme in Experimental Biology and Biomedicine of the Center for Neuroscience and Cell Biology, University of Coimbra (PD/BD/129294/2017). Work in AE and MM labs on microRNAs and infection is supported by the ERA-NET Infect-ERA CampyRNA and the FCT R&D grant (#POCI-01-0145-FEDER-029999).
Publisher Copyright:
© 2018 Elsevier Inc.
PY - 2019/1/1
Y1 - 2019/1/1
N2 - MicroRNAs (miRNAs) are small non-coding RNAs that post-transcriptionally modulate gene expression and orchestrate a wide range of biological and pathological processes. The use of high-throughput screening technologies, in particular microscopy-based screenings (also known as high-content screenings), coupled with genome-wide libraries for modulation of miRNA levels, allow for comprehensive functional analysis of each member of the miRNome in different phenotypic cell-based assays. The wealth of information obtained from such screenings spans across various fields of research, including cancer, cardiovascular, cell reprogramming, and infection biology. Here, we provide an overview of the rationale for performing screenings using synthetic libraries of miRNA mimics and inhibitors, and of the microscopy-based miRNA screenings performed to date. Moreover, a list of resources available for such endeavor is provided. Finally, we describe a detailed procedure for a case study where microscopy-based screening using a library of miRNA mimics was performed to identify miRNAs that control infection of epithelial cells by the bacterial pathogen Salmonella. The methodologies described here can be easily adapted for screenings addressing other biological questions.
AB - MicroRNAs (miRNAs) are small non-coding RNAs that post-transcriptionally modulate gene expression and orchestrate a wide range of biological and pathological processes. The use of high-throughput screening technologies, in particular microscopy-based screenings (also known as high-content screenings), coupled with genome-wide libraries for modulation of miRNA levels, allow for comprehensive functional analysis of each member of the miRNome in different phenotypic cell-based assays. The wealth of information obtained from such screenings spans across various fields of research, including cancer, cardiovascular, cell reprogramming, and infection biology. Here, we provide an overview of the rationale for performing screenings using synthetic libraries of miRNA mimics and inhibitors, and of the microscopy-based miRNA screenings performed to date. Moreover, a list of resources available for such endeavor is provided. Finally, we describe a detailed procedure for a case study where microscopy-based screening using a library of miRNA mimics was performed to identify miRNAs that control infection of epithelial cells by the bacterial pathogen Salmonella. The methodologies described here can be easily adapted for screenings addressing other biological questions.
KW - Bacterial infection
KW - High-content microscopy
KW - MicroRNA
KW - MicroRNA libraries
KW - Microscopy-based screening
KW - Salmonella
UR - http://www.scopus.com/inward/record.url?scp=85054618739&partnerID=8YFLogxK
U2 - 10.1016/j.ymeth.2018.09.011
DO - 10.1016/j.ymeth.2018.09.011
M3 - Review article
C2 - 30292796
AN - SCOPUS:85054618739
SN - 1046-2023
VL - 152
SP - 55
EP - 64
JO - Methods
JF - Methods
ER -
