TY - JOUR
T1 - Beyond quantification
T2 - in situ analysis of transcriptome and pre-mRNA alternative splicing at the nanoscale
AU - Cui, Yi
AU - Liu, Jing
AU - Irudayaraj, Joseph
N1 - Publisher Copyright:
© 2016 Wiley Periodicals, Inc.
PY - 2017/7/1
Y1 - 2017/7/1
N2 - In situ analysis offers a venue for dissecting the complex transcriptome in its natural context to tap into cellular processes that could explain the phenotypic physiology and pathology yet to be understood. Over the past decades, enormous progress has been made to improve the resolution, sensitivity, and specificity of single-cell technologies. The continued efforts in RNA research not only facilitates mechanistic studies of molecular biology but also provides state-of-the-art strategies for diagnostic purposes. The implementation of novel bio-imaging platforms has yielded valuable information for inspecting gene expression, mapping regulatory networks, and classifying cell types. In this article, we discuss the merits and technical challenges in single-molecule in situ RNA profiling. Advanced in situ hybridization methodologies developed for a variety of detection modalities are reviewed. Considering the fact that in mammalian cells the number of protein products immensely exceeds that of the actual coding genes due to pre-mRNA alternative splicing, tools capable of elucidating this process in intact cells are highlighted. To conclude, we point out future directions for in situ transcriptome analysis and expect a plethora of opportunities and discoveries in this field. WIREs Nanomed Nanobiotechnol 2017, 9:e1443. doi: 10.1002/wnan.1443. For further resources related to this article, please visit the WIREs website.
AB - In situ analysis offers a venue for dissecting the complex transcriptome in its natural context to tap into cellular processes that could explain the phenotypic physiology and pathology yet to be understood. Over the past decades, enormous progress has been made to improve the resolution, sensitivity, and specificity of single-cell technologies. The continued efforts in RNA research not only facilitates mechanistic studies of molecular biology but also provides state-of-the-art strategies for diagnostic purposes. The implementation of novel bio-imaging platforms has yielded valuable information for inspecting gene expression, mapping regulatory networks, and classifying cell types. In this article, we discuss the merits and technical challenges in single-molecule in situ RNA profiling. Advanced in situ hybridization methodologies developed for a variety of detection modalities are reviewed. Considering the fact that in mammalian cells the number of protein products immensely exceeds that of the actual coding genes due to pre-mRNA alternative splicing, tools capable of elucidating this process in intact cells are highlighted. To conclude, we point out future directions for in situ transcriptome analysis and expect a plethora of opportunities and discoveries in this field. WIREs Nanomed Nanobiotechnol 2017, 9:e1443. doi: 10.1002/wnan.1443. For further resources related to this article, please visit the WIREs website.
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U2 - 10.1002/wnan.1443
DO - 10.1002/wnan.1443
M3 - Review article
C2 - 27813271
AN - SCOPUS:84996538656
SN - 1939-5116
VL - 9
JO - Wiley Interdisciplinary Reviews: Nanomedicine and Nanobiotechnology
JF - Wiley Interdisciplinary Reviews: Nanomedicine and Nanobiotechnology
IS - 4
M1 - e1443
ER -