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Method Article
Zhiguang Guo
Guo's Lab, Sanford Research, 2301 East 60th Street North, Sioux Falls, SD 57104, USA
Corresponding Author
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Pancreatic islets primarily involved in regulating blood glucose homeostasis are the key focus of diabetes study and miRNA-mediated molecular regulation. Whole mount in situ hybridization of miRNAs and co-localization of proteins is a powerful tool for studying cell-specific expression and regulation. Although many miRNAs are potential regulators of islet function and diabetes development, most of their cell specific expression remains unknown. We developed and optimized a protocol that combines fluorescent in situ hybridization (FISH) of miRNAs and immunofluorescence (IF) of two proteins at the same time in whole mount isolated intact islets for preserving islet morphology, eliminating the technical difficulties associated with handling isolated islets that are not amenable to conventional FISH, as well as for increasing sensitivity and resolution. This method can serve as a valuable tool for studying miRNA-mediated regulation of post-transcriptional gene expression in islet biology, diabetes research, and biomarker development.
Keywords
Fluorescent In situ Hybridization, miRNA, Immunofluorescence, Islets
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The authors declare no competing financial interests.
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Associated Publications
Identification and Characterization of microRNAs Associated With Human β-Cell Loss in a Mouse Model
by R. Roat, M. M. Hossain, J. Christopherson, +5
Method Article
Zhiguang Guo
Guo's Lab, Sanford Research, 2301 East 60th Street North, Sioux Falls, SD 57104, USA
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 12 Apr, 2018
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Pancreatic islets primarily involved in regulating blood glucose homeostasis are the key focus of diabetes study and miRNA-mediated molecular regulation. Whole mount in situ hybridization of miRNAs and co-localization of proteins is a powerful tool for studying cell-specific expression and regulation. Although many miRNAs are potential regulators of islet function and diabetes development, most of their cell specific expression remains unknown. We developed and optimized a protocol that combines fluorescent in situ hybridization (FISH) of miRNAs and immunofluorescence (IF) of two proteins at the same time in whole mount isolated intact islets for preserving islet morphology, eliminating the technical difficulties associated with handling isolated islets that are not amenable to conventional FISH, as well as for increasing sensitivity and resolution. This method can serve as a valuable tool for studying miRNA-mediated regulation of post-transcriptional gene expression in islet biology, diabetes research, and biomarker development.
Figure 1
Figure 2
Figure 3
Figure 4
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Associated Publications
Identification and Characterization of microRNAs Associated With Human β-Cell Loss in a Mouse Model
by R. Roat, M. M. Hossain, J. Christopherson, +5