In vivo methods for acute modulation of gene expression in the central nervous system

Abstract

Accurate and timely expression of specific genes guarantees the healthy development and function of the brain. Indeed, variations in the correct amount or timing of gene expression lead to improper development and/or pathological conditions. Almost forty years after the first successful gene transfection in in vitro cell cultures, it is currently possible to regulate gene expression in an area-specific manner at any step of central nervous system development and in adulthood in experimental animals in vivo, even overcoming the very poor accessibility of the brain. Here, we will review the diverse approaches for acute gene transfer in vivo, highlighting their advantages and disadvantages with respect to the efficiency and specificity of transfection as well as to brain accessibility. In particular, we will present well-established chemical, physical and virus-based approaches suitable for different animal models, pointing out their current and future possible applications in basic and translational research as well as in gene therapy.

Niosome-Based Approach for In Situ Gene Delivery to Retina and Brain Cortex as Immune-Privileged Tissues

“…Although highly effective, these methods are normally restricted to ex vivo gene therapy, which is mainly due to the challenge that represents the fine control of physical parameters that produce the formation of transient pores into the cellular membrane in vivo conditions. Such technological limitations can result not only in a loss of action but also in an increase of the cellular toxicity [45]. Transient pores on cellular and even nuclear membranes can be induced by the application of external electrical pulses, whose amplitude and duration are controlled, and depend on the particular characteristics of the target cell [22].…”

Battling Neurodegenerative Diseases with Adeno-Associated Virus-Based Approaches

“…Depending on the nature of target cells, gene delivery can be classified as either somatic or germline. While the goal of somatic gene delivery is to directly express the transgene in target cells and correct the disease at the individual cell level [39], germline gene transfer is aimed at integrating a therapeutic or correction gene (fully functional wild type copy) into the germline cells, thereby creating inheritable changes, affecting every cell of the next generation. Although more advanced techniques of gene delivery are constantly being developed, the most common approach to correct NDDs involves a direct transfection or transduction (when using virus-based delivery vectors) of isolated patient-derived cells or modification of target cells in situ-i.e., directly in patient's tissue.…”

Identification and prevention of heterotopias in mouse neocortical neural cell migration incurred by surgical damages during utero electroporation procedures

“…The expression vectors are microinjected into the embryonic brain ventricle, and transfect the neuronal precursor cells with the DNA by squarewave electric pulses (Niwa et al 2010;Youngpearse et al 2010). Using this technique, the DNA can be delivered or transfected into the cerebral cortex, diencephalon, hindbrain, and spinal cord region (David et al 2014;Nicole et al 2018;Watanabe et al 2018) and used for the evaluation of the phenotypic changes that are relevant to the disease pathology (Cwetsch et al 2018). But IUE technique has different drawbacks that could lead to different unexpected false or imitation results, such as surgical damages to cause artificial ectopias and heterotopias.…”

“…The major disadvantages of these methods are such as limitation in transfection efficiency and specio-temporal gene manipulation, anduncontrolled or non-specified transfection. IUE has several advantages over the traditional techniques like higher transfection efficiency, lower cytotoxicity, time-efficient, and multiple plasmids can be simultaneously transfected into the same cells (Saito 2006;Cwetsch et al 2018). IUE requires neither complex surgery nor stereotaxic apparatus, so now, IUE is widely used to understand the mechanism of brain development (Carrel et al 2015;Broix et al 2018).…”