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A critical analysis of methods used to investigate the cellular uptake and subcellular localization of RNA therapeutics
- Source :
- Nucleic Acids Research
- Publication Year :
- 2020
- Publisher :
- Oxford University Press (OUP), 2020.
-
Abstract
- RNA therapeutics are a promising strategy to treat genetic diseases caused by the overexpression or aberrant splicing of a specific protein. The field has seen major strides in the clinical efficacy of this class of molecules, largely due to chemical modifications and delivery strategies that improve nuclease resistance and enhance cell penetration. However, a major obstacle in the development of RNA therapeutics continues to be the imprecise, difficult, and often problematic nature of most methods used to measure cell penetration. Here, we review these methods and clearly distinguish between those that measure total cellular uptake of RNA therapeutics, which includes both productive and non-productive uptake, and those that measure cytosolic/nuclear penetration, which represents only productive uptake. We critically analyze the benefits and drawbacks of each method. Finally, we use key examples to illustrate how, despite rigorous experimentation and proper controls, our understanding of the mechanism of gymnotic uptake of RNA therapeutics remains limited by the methods commonly used to analyze RNA delivery.
- Subjects :
- Specific protein
AcademicSubjects/SCI00010
Computational biology
Biology
010402 general chemistry
01 natural sciences
03 medical and health sciences
Cytosol
Genetics
Humans
Clinical efficacy
Survey and Summary
RNA, Small Interfering
Aberrant splicing
030304 developmental biology
Cell Nucleus
RNA metabolism
0303 health sciences
Oligonucleotide
Genetic Diseases, Inborn
RNA
Aptamers, Nucleotide
Oligonucleotides, Antisense
Subcellular localization
0104 chemical sciences
MicroRNAs
Microscopy, Electron
Spectrometry, Fluorescence
Genetic Techniques
Cell penetration
Subjects
Details
- ISSN :
- 13624962 and 03051048
- Volume :
- 48
- Database :
- OpenAIRE
- Journal :
- Nucleic Acids Research
- Accession number :
- edsair.doi.dedup.....155b472acc5745a6ecbe3df8ec92b162