Chapter title |
Design of shRNA and miRNA for Delivery to the CNS.
|
---|---|
Chapter number | 5 |
Book title |
Gene Therapy for Neurological Disorders
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3271-9_5 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3270-2, 978-1-4939-3271-9
|
Authors |
Toro Cabrera, Gabriela, Mueller, Christian, Gabriela Toro Cabrera, Christian Mueller |
Abstract |
Neurologic diseases tend to target various areas of the central nervous system (CNS) and can therefore result in paralysis, dementia, and death. Neurodegenerative diseases distinguish themselves from other diseases by affecting nerve cells, which unlike many other cells in our body cannot regenerate when severely injured. The discovery of RNA interference (RNAi) has enabled scientist to design new therapeutic approaches based on specific gene silencing rather than the canonical gene therapy through gene augmentation. Two types of molecules can be used for viral vector-mediated gene silencing: short hairpin RNAs (shRNAs) and artificial microRNAs (miRNAs) that have the ability to enter the RNAi pathway. Although both shRNAs and miRNAs can be used to silence genes, they enter the RNAi pathway at different points. Unlike shRNAs, miRNAs require an additional cleavage step inside the nucleus before being exported to the cytoplasm. These molecules can then be incorporated into the RNA-induced silencing complex (RISC) which utilizes sequence complementarity to recognize target mRNAs and activate either translational repression, in the case of partial complementarity, or induce mRNA cleavage in the case of complete complementarity. Elevated amounts of shRNAs, which are commonly driven by strong polymerase III promoters, can cause saturation of the endogenous RNAi machinery due to competition between endogenous and artificial molecules. Switching to a DNA polymerase II promoter is an alternative to reduce shRNA production, thereby reducing toxicity. Even though the molecules are designed to target specific mRNAs there may be off-target effects due to nonspecific binding that must be accounted for during the design process. In this chapter we discuss the design and in vitro screening of shRNAs and artificial miRNAs. |
X Demographics
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United States | 1 | 25% |
Unknown | 2 | 50% |
Demographic breakdown
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Members of the public | 2 | 50% |
Scientists | 1 | 25% |
Science communicators (journalists, bloggers, editors) | 1 | 25% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
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Unknown | 26 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Researcher | 7 | 27% |
Student > Ph. D. Student | 6 | 23% |
Student > Bachelor | 3 | 12% |
Student > Doctoral Student | 2 | 8% |
Other | 2 | 8% |
Other | 4 | 15% |
Unknown | 2 | 8% |
Readers by discipline | Count | As % |
---|---|---|
Neuroscience | 7 | 27% |
Biochemistry, Genetics and Molecular Biology | 6 | 23% |
Agricultural and Biological Sciences | 5 | 19% |
Medicine and Dentistry | 2 | 8% |
Veterinary Science and Veterinary Medicine | 1 | 4% |
Other | 1 | 4% |
Unknown | 4 | 15% |