Abstract
Molecular beacons (MBs) are synthetic oligonucleotide probes that are designed to fluoresce upon hybridization to complementary nucleic acid targets. In contrast to genetically encoded probes that can be readily introduced into cells via standard transfection procedures, using MBs to obtain reliable intracellular measurements entails a reliable delivery method that maximizes MB entry while minimizing cell damage. One promising approach is microporation, a microliter volume electroporation-based method that exhibits reduced harmful events as compared with traditional electroporation methods. In this chapter, we describe in detail microporation steps for MB delivery that we have utilized over the past several years, followed by examples demonstrating successful MB-based imaging of specific RNA transcripts and genomic loci at the single-molecule level.
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Acknowledgments
This project was supported by grants from the National Key R&D Program of China (No. 2016YFA0501603), the National Natural Science Foundation of China (No. 31771583), and the Beijing Municipal R&D Key Project (Grant No. Z151100003915081).
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Mao, S., Ying, Y., Wu, X., Chen, A.K. (2020). Delivering Molecular Beacons via an Electroporation-Based Approach Enables Live-Cell Imaging of Single RNA Transcripts and Genomic Loci. In: Heise, T. (eds) RNA Chaperones. Methods in Molecular Biology, vol 2106. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0231-7_15
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DOI: https://doi.org/10.1007/978-1-0716-0231-7_15
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