Chapter title |
Non-Viral, Lipid-Mediated DNA and mRNA Gene Therapy of the Central Nervous System (CNS): Chemical-Based Transfection
|
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Chapter number | 23 |
Book title |
Gene Therapy for Neurological Disorders
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Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3271-9_23 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3270-2, 978-1-4939-3271-9
|
Authors |
James G. Hecker, Hecker, James G. |
Abstract |
Appropriate gene delivery systems are essential for successful gene therapy in clinical medicine. Cationic lipid-mediated delivery is an alternative to viral vector-mediated gene delivery. Lipid-mediated delivery of DNA or mRNA is usually more rapid than viral-mediated delivery, offers a larger payload, and has a nearly zero risk of incorporation. Lipid-mediated delivery of DNA or RNA is therefore preferable to viral DNA delivery in those clinical applications that do not require long-term expression for chronic conditions. Delivery of RNA may be preferable to non-viral DNA delivery in some clinical applications, because transit across the nuclear membrane is not necessary and onset of expression with RNA is therefore even faster than with DNA, although both are faster than most viral vectors. Here, we describe techniques for cationic lipid-mediated delivery of nucleic acids encoding reporter genes in a variety of cell lines. We describe optimized formulations and transfection procedures that we previously assessed by bioluminescence and flow cytometry. RNA transfection demonstrates increased efficiency relative to DNA transfection in non-dividing cells. Delivery of mRNA results in onset of expression within 1 h after transfection and a peak in expression 5-7 h after transfection. Duration of expression in eukaryotic cells after mRNA transcript delivery depends on multiple factors, including transcript stability, protein turnover, and cell type. Delivery of DNA results in onset of expression within 5 h after transfection, a peak in expression 24-48 h after transfection, and a return to baseline that can be as long as several weeks after transfection. In vitro results are consistent with our in vivo delivery results, techniques for which are described as well. RNA delivery is suitable for short-term transient gene expression due to its rapid onset, short duration of expression and greater efficiency, particularly in non-dividing cells, while the longer duration and the higher mean levels of expression per cell that are ultimately obtained following DNA delivery confirm a continuing role for DNA gene delivery in clinical applications that require longer term transient gene expression. |
X Demographics
Geographical breakdown
Country | Count | As % |
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United States | 1 | 14% |
Unknown | 6 | 86% |
Demographic breakdown
Type | Count | As % |
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Members of the public | 6 | 86% |
Practitioners (doctors, other healthcare professionals) | 1 | 14% |
Mendeley readers
Geographical breakdown
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Spain | 1 | 3% |
Unknown | 33 | 97% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Master | 10 | 29% |
Student > Ph. D. Student | 6 | 18% |
Student > Bachelor | 3 | 9% |
Researcher | 3 | 9% |
Other | 2 | 6% |
Other | 1 | 3% |
Unknown | 9 | 26% |
Readers by discipline | Count | As % |
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Biochemistry, Genetics and Molecular Biology | 10 | 29% |
Pharmacology, Toxicology and Pharmaceutical Science | 8 | 24% |
Agricultural and Biological Sciences | 3 | 9% |
Nursing and Health Professions | 1 | 3% |
Immunology and Microbiology | 1 | 3% |
Other | 3 | 9% |
Unknown | 8 | 24% |