Methods for Gene Transfer to the Central Nervous System

Abstract

Gene transfer is an increasingly utilized approach for research and clinical applications involving the central nervous system (CNS). Vectors for gene transfer can be as simple as an unmodified plasmid, but more commonly involve complex modifications to viruses to make them suitable gene delivery vehicles. This chapter will explain how tools for CNS gene transfer have been derived from naturally occurring viruses. The current capabilities of plasmid, retroviral, adeno-associated virus, adenovirus, and herpes simplex virus vectors for CNS gene delivery will be described. These include both focal and global CNS gene transfer strategies, with short- or long-term gene expression. As is described in this chapter, an important aspect of any vector is the cis-acting regulatory elements incorporated into the vector genome that control when, where, and how the transgene is expressed.

A Protocol for the Production of Integrase-deficient Lentiviral Vectors for CRISPR/Cas9-mediated Gene Knockout in Dividing Cells

“…These advantages make IDLVs a powerful tool in basic science and clinical research (reviewed in Kantor et al, 6 and Wanisch and Yáñez-Muñoz 22 ). 24,58 Throughout this study, we sought to establish a single-molecule IDLV platform for efficient delivery of CRISPR/Cas9 components in vitro and in vivo.…”

“…20 Nevertheless, the packaging efficacy of all-in-one AAV vector systems, especially those intended for clinical applications, needs further improvement to efficiently accommodate multiple components, including the 3.2-kb SaCas9, the RNA polymerase II (Pol II) promoter and poly(A), a nuclear localization signal (NLS), sgRNA(s), and the RNA polymerase III (Pol III) promoter(s), as well as other cis-acting elements, such as woodchuck hepatitis post-transcriptional regulatory element (WPRE). 21 Episomal IDLVs are an ideal platform for delivery of large genetic cargos where only transient expression of the transgene is desired (reviewed in Kantor et al, 6 and Wanisch and Yáñez-Muñoz 22 ).…”

“…[3][4][5] Lentiviral vectors (LVs) are an important means of delivering CRISPR/Cas9 components due to their ability to accommodate large DNA payloads and efficiently transduce a wide range of dividing and non-dividing cells. LVs also display low cytotoxicity and immunogenicity and have a minimal impact on the life cycle of the transduced cells (reviewed in Kantor et al 6 ). Such features have led to LVs being used as the gene-editing regimen of choice to treat HIV-1, HBV, and HSV-1 infections, as well as to correct defects underlying human hereditary diseases, such as cystic fibrosis.…”

Selective Manipulation of Neural Circuits

“…Now, several serotypes of adeno-associated virus (AAV) such as AAV6, AAV8, and AAV9 are characterized to transduce neurons retrogradely [61][62][63][64][65], and lentivirus has been modified with its glycoprotein fused with rabies virus glycoprotein for retrograde transduction [41,42,66,67]. These viruses are easy to produce, result in less inflammation than herpes simplex and rabiesderived vectors, and have long duration and high level of transgene expression [68][69][70][71][72].…”

Nanoparticle-Mediated Upconversion of Near-Infrared Light: A Step Closer to Optogenetic Neuromodulation in Humans

“…Optogenetics has been successfully used to treat Parkinson’s disease, chronic pain, refractory epilepsy and retinitis pigmentosa in different animal models [2]. However, therapeutic deep brain optogenetic neuromodulation in humans is limited by a number of practical challenges, notably the long-term safety of viral vectors used for genetic engineering [8-11], the long-term expression of light-reactive channels in neurons [12], the effective delivery and diffusion of light through brain tissue [13, 14], tissue damage by light-related overheating [13, 14] and the still rudimentary understanding of network dynamics in the human brain [7, 8, 15]. …”