Using viral vectors as gene transfer tools (Cell Biology and Toxicology Special Issue: ETCS-UK 1 day meeting on genetic manipulation of cells)

Abstract

In recent years, the development of powerful viral gene transfer techniques has greatly facilitated the study of gene function. This review summarises some of the viral delivery systems routinely used to mediate gene transfer into cell lines, primary cell cultures and in whole animal models. The systems described were originally discussed at a 1-day European Tissue Culture Society (ETCS-UK) workshop that was held at University College London on 1st April 2009. Recombinant-deficient viral vectors (viruses that are no longer able to replicate) are used to transduce dividing and post-mitotic cells, and they have been optimised to mediate regulatable, powerful, long-term and cell-specific expression. Hence, viral systems have become very widely used, especially in the field of neurobiology. This review introduces the main categories of viral vectors, focusing on their initial development and highlighting modifications and improvements made since their introduction. In particular, the use of specific promoters to restrict expression, translational enhancers and regulatory elements to boost expression from a single virion and the development of regulatable systems is described.

Development of high-concentration lipoplexes for in vivo gene function studies in vertebrate embryos

“…Enters cells very efficiently including in post-mitotic cells (Brugmann et al, 2007;Howarth et al, 2010). All cationic lipds are similar, they contain a positive headgroup and a hydrophobic tail .…”

Viral Vectors in Neurobiology: Therapeutic and Research Applications

“…Otherwise, non-integrative vectors, like adenovirus whose viral DNA is maintained in an episomal form into infected cells, lead to a transient transgene expression. Each vector presents specific advantages and limitations that become more or less proper depending on the objective of its application (http://cmbi.bjmu.edu.cn/cmbidata/therapy/research/re02/021.htm; Osten et al, 2007;Howarth et al, 2010).…”

“…On the other hand, these vectors show poor in vivo delivery and present the risk of insertional mutagenesis. However, as lentiviruses mostly transduce terminally differentiated cells, the risk of insertional mutagenesis is less important than observed for other retroviruses (Kaplitt & Pfaff, 1996;Davidson & Breakefield, 2003;Howarth et al, 2010;Snyder et al, 2010).…”

“…The adult brain is composed largely of non-dividing cells, and therefore DNA viruses have become the vehicle of choice for neurobiologists interested in somatic gene transfer. By re-expressing an absent protein or by overexpressing a certain protein (i.e increasing its concentration), one can unravel its functions and possible process that it underlies (Howarth et al, 2010).…”

Novel approaches to in vitro transgenesis

“…A number of viral-based vectors are commonly used to ectopically express reporter constructs, including adenovirus (AV), adeno-associated virus (AAV), herpes simplex virus (HSV) and retroviruses (principally lentiviruses (LV)). Each viral vector has a number of advantages and disadvantages and this has been reviewed extensively elsewhere (Verma & Weitzman 2005, Osten et al 2007, Howarth et al 2010. In this review, we present a brief summary of the general properties of viral transduction and summarise the characteristics and properties of commonly used viral vectors (Table 1).…”