Adeno-Associated Virus Technologies and Methods for Targeted Neuronal Manipulation

Abstract

Cell-type-specific expression of molecular tools and sensors is critical to construct circuit diagrams and to investigate the activity and function of neurons within the nervous system. Strategies for targeted manipulation include combinations of classical genetic tools such as Cre/loxP and Flp/FRT, use of cis-regulatory elements, targeted knock-in transgenic mice, and gene delivery by AAV and other viral vectors. The combination of these complex technologies with the goal of precise neuronal targeting is a challenge in the lab. This report will discuss the theoretical and practical aspects of combining current technologies and establish best practices for achieving targeted manipulation of specific cell types. Novel applications and tools, as well as areas for development, will be envisioned and discussed.Selecting the Route of Administration and Capsid AAV tropism, as dictated by AAV capsid proteins, is an important factor affecting transduction efficiency and specificity across cell types. Since the mechanism of AAV transduction is through interaction of the AAV capsid with cell surface proteins and glycans, protein composition of the capsid (i.e., the AAV serotype) and the cell surface (i.e., based on cell type) determines transduction efficiency. Consequently, serotype and route of delivery should be carefully considered when designing experiments (Fig 1A, B). For an overview of the primary receptors for AAV serotypes, see (Schultz and Chamberlain, 2008). Direct intraparenchymal deliveryWhen injected directly into the brain, many of the naturally-occurring AAV capsids, which share homology ranging from 65-99% (Drouin and Agbandje-McKenna, 2013), have distinct but significantly overlapping tropisms and distribution characteristics. AAV1, AAV2, AAV5, AAV8,

Application of Recombinant Rabies Virus toXenopusTadpole Brain

“…Virus-mediated gene expression in non-mammalian vertebrates has had mixed results. AAV and lentivirus, which have been used with great success in mammals (Haery et al, 2019;Nassi et al, 2015;Parr-Brownlie et al, 2015), infect Xenopus and zebrafish either inconsistently or not at all (Yamaguchi et al, 2018;Zhu et al, 2009). Xenopus express two homologues of the Cosackie and Adenovirus Receptor (CAR) rendering them susceptible to infection by adenovirus and adenovirus-mediated expression of EGFP lasts for at least 10 days in non-neuronal tissue (Dutton et al, 2009;Kawakami et al, 2006).…”

Histaminergic neurons in the tuberomammillary nucleus as a control centre for wakefulness

“…Adeno-associated virus are classified into 10 different serotypes which exhibit differences in transduction efficacy, spread from injection site and directional characteristics (e.g. neuron-specific transduction or neuronal/glial transduction) (Haery et al, 2019).…”

New roles for dopamine in motor skill acquisition: lessons from primates, rodents, and songbirds

“…Mice in particular offer a level of specificity in 129 neural circuit manipulation and labeling that is unparalleled among mammals due to the wide array of sophisticated and accessible molecular genetic tools (e.g. [40][41][42]. While molecular tools in both nonhuman primates and birds are developing rapidly (43,44), there is no comparison at this point with the wide range of tools available in mice.…”