Efficient CNS gene delivery by intravenous injection. Nat Methods 7:905-907

Department of Pathology, Thomas Jefferson University, Philadelphia, Pennsylvania, USA.
Nature Methods (Impact Factor: 32.07). 11/2010; 7(11):905-7. DOI: 10.1038/nmeth.1518
Source: PubMed


We administered recombinant SV40-derived viral vectors (rSV40s) intravenously to mice with or without prior intraperitoneal injection of mannitol to deliver transgenes to the central nervous system (CNS). We detected transgene-expressing cells (mainly neurons) most prominently in the cortex and spinal cord; prior intraperitoneal mannitol injection increased CNS gene delivery tenfold. Intravenous injection of rSV40s, particularly with mannitol pretreatment, resulted in extensive expression of multiple transgenes throughout the CNS.

Download full-text


Available from: Jayanta Roy-Chowdhury, Mar 24, 2014
  • Source
    • "The alternative methods, including comparison of neural injury scores or quantification of the area devoid of Nissl or MAP2 staining, are all either semi-quantitative or labor-intensive. Hence, inspired by recent reports of mannitolfacilitated CNS-entry of viral vectors (McCarty et al., 2009; Louboutin et al., 2010), we adapted this appraoch to develop a TTC perfusion-labeling method. The new in-vivo TTClabeling method is simple, fast, and reliable. "
    [Show abstract] [Hide abstract]
    ABSTRACT: A simple method to quantify cerebral infarction has great value for mechanistic and therapeutic studies in experimental stroke research. Immersion staining of unfixed brain slices with 2,3,5-triphenyltetrazolium chloride (TTC) is a popular method to determine cerebral infarction in preclinical studies. However, it is often difficult to apply immersion TTC-labeling to severely injured or soft newborn brains in rodents. Here we report an in vivo TTC perfusion-labeling method based on osmotic opening of blood-brain-barrier with mannitol-pretreatment. This new method delineates cortical infarction correlated with the boundary of morphological cell injury, differentiates the induction or subcellular redistribution of apoptosis-related factors between viable and damaged areas, and easily determines the size of cerebral infarction in both adult and newborn mice. Using this method, we confirmed that administration of lipopolysaccharide 72 h before hypoxia-ischemia increases the damage in neonatal mouse brains, in contrast to its effect of protective preconditioning in adults. These results demonstrate a fast and inexpensive method that simplifies the task of quantifying cerebral infarction in small or severely injured brains and assists biochemical analysis of experimental cerebral ischemia.
    Full-text · Article · Jan 2012 · Journal of Neuroscience Methods
  • Source
    • "Recently, two types of viral vectors were found to cross the BBB. The adenoassociated virus type 9 (AAV9) vector was found to cross the BBB of neonate mice, cats, as well as nonhuman primates after intravenous injection [5-7], and simian virus 40 (SV40) was able to cross the BBB of mice [8]. Nevertheless, further studies are needed to check if this feature remains in other mammalian species, including humans. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Alzheimer's and Parkinson's diseases represent the most prevalent neurodegenerative disorders worldwide. Current pharmacological or surgical treatments provide symptomatic benefits, particularly in the early stages, but none can delay or stop the progression of these diseases. There is an urgent need for new therapies able to modify disease progression. Gene therapy, mainly based on viral vectors, is presently being assumed as an important alternative to conventional treatments. After decades of preclinical developments, we are now facing an important period, in which several viral vector-mediated gene therapies are being tested in Phase 1 and Phase 2 clinical trials, with some of them showing promising results. This review intends to present an overview of the current efforts in the field for the treatment of Alzheimer's and Parkinson's diseases.
    Full-text · Article · Sep 2011 · Current pharmaceutical design
  • [Show abstract] [Hide abstract]
    ABSTRACT: Endogenous RNA-silencing mechanisms have been shown to play a role in regulating viral and host processes during the course of infection. Such interactive processes may involve host cellular and/or viral-encoded microRNAs (miRNAs). Rabies is unique not only in terms of its invariably fatal course once disease signs develop, but it also has a variable incubation period (eclipse phase). It has been recently shown that cells or tissues of different origin have their own specific miRNAs that, in theory, may impact on viral transcription and replication. This may possibly explain, in part, why rabies virus remains dormant at the inoculation site in rabies patients for long periods. Owing to the RNA interference (RNAi) technology, it has been possible to introduce exogenously designed artificial short interfering RNAs (siRNAs) and miRNAs into virus-infected cells for therapeutic purposes. Successful attempts in using RNAi for prevention and treatment of DNA and RNA virus infections both in vitro and in vivo experiments have been reported. The fact that rabies remains incurable has stimulated the development of the therapeutic RNAi strategy. We describe herein preliminary evidence that cellular miRNA may play a role in suppressing viral replication, explaining the eclipse phase, and that artificially designed multitargeting miRNA can successfully inhibit rabies virus transcription and replication in vitro.
    No preview · Article · Dec 2011 · Advances in Virus Research
Show more