Delivery Platforms for CRISPR/Cas9 Genome Editing of Glial Cells in the Central Nervous System

Abstract

Glial cells (astrocytes, oligodendrocytes, and microglia) are emerging as key players in several physiological and pathological processes of the central nervous system (CNS). Astrocytes and oligodendrocytes are not only supportive cells that release trophic factors or regulate energy metabolism, but they also actively modulate critical neuronal processes and functions in the tripartite synapse. Microglia are defined as CNS-resident cells that provide immune surveillance; however, they also actively contribute to shaping the neuronal microenvironment by scavenging cell debris or regulating synaptogenesis and pruning. Given the many interconnected processes coordinated by glial cells, it is not surprising that both acute and chronic CNS insults not only cause neuronal damage but also trigger complex multifaceted responses, including neuroinflammation, which can critically contribute to the disease progression and worsening of symptoms in several neurodegenerative diseases. Overall, this makes glial cells excellent candidates for targeted therapies to treat CNS disorders. In recent years, the application of gene editing technologies has redefined therapeutic strategies to treat genetic and age-related neurological diseases. In this review, we discuss the advantages and limitations of clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9-based gene editing in the treatment of neurodegenerative disorders, focusing on the development of viral- and nanoparticle-based delivery methods for in vivo glial cell targeting.

GABAA Receptors in Astrocytes Are Targets for Commonly Used Intravenous and Inhalational General Anesthetic Drugs

“…Future studies will determine whether astrocytic GABA A receptors contribute to this crosstalk with neuronal GABA A receptors that may contribute to PNDs, as well as identify the soluble factor(s). Using genetic approaches, such as astrocyte-specific gene knockdown/knockout, and novel CRISPR-based technology to target anesthetic-sensitive astrocytic GABA A receptors in vitro and in vivo , may help answer these questions ( Mori et al, 2006 ; Shinohara et al, 2016 ; Meneghini et al, 2021 ). Such studies may lead to the discovery of novel strategies to mitigate the cognitive dysfunction experienced by older patients with PNDs.…”

Scientific Evidences of Calorie Restriction and Intermittent Fasting for Neuroprotection in Traumatic Brain Injury Animal Models: A Review of the Literature

“…It is generally accepted that glial cell activation following brain injury plays a critical role in the progression of secondary injury following the primary insult [ 70 , 71 , 72 , 73 , 74 ], such as triggering the neuroinflammatory responses [ 75 , 76 , 77 , 78 , 79 ]. Adult male mice on a three-month CR regimen (50% of a normal daily diet) prior to TBI were found to have significantly reduced microglia activation one month after injury.…”

Recent advances in the use of CRISPR/Cas for understanding the early development of molecular gaps in glial cells

“…In addition, differentiation employing small molecules has shown better epigenome reprogramming which is essential for reliable iPSC-based models (Meneghini et al, 2021;Guan et al, 2022).…”

“…Additionally, the NHEJ pathway is more active and error-prone, facilitating frameshift mutations in the coding sequence. Although new approaches have been developed such as the strategy "HiTi" (homology-independent targeted integration) to improve DNA knock-in in dividing and nondividing cells (Vesikansa, 2018;Meneghini et al, 2021).…”