Neuron-Specific Transduction in the Rat Septohippocampal or Nigrostriatal Pathway by Recombinant Adeno-associated Virus Vectors

Klein, Ronald L.; Meyer, Edwin M.; Peel, Alyson; Zolotukhin, Sergei; Meyers, Craig; Muzyczka, Nicholas; King, Michael A.

doi:10.1006/exnr.1997.6736

Cited by 229 publications

(155 citation statements)

References 41 publications

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“…Under optimal conditions, as few as 10-40 infectious particles of rAAV have been found to be sufficient to transduce one cell in vivo. 6,7,9 During the course of these studies at least three problems were routinely encountered during purification of rAAV using conventional cesium chloride centrifugation methods. First, rAAV is often nonspecifically bound to cellular protein and helper Ad virus.…”

Section: Discussionmentioning

confidence: 99%

“…In Figure 3, however, the gradient was collected by dripping from the bottom and collecting 1 ml fractions for detailed analysis. The bulk of the rAAV virus bands within the 40% density step (Figure 3c, fractions [5][6][7][8]. A heavy band that migrates at the 40%-to-25% density junction consists mostly of cellular proteins (Figure 3c, fractions 9 and 10) and contains less then 5% input rAAV, as judged by FCA.…”

Section: Figure 2 Titering Of Raav Stocks By the Qc Pcr Assay (A) Or mentioning

confidence: 96%

“…6 To produce the enhanced blue fluorescent mutant 37 of gfp, we introduced the Tyr145Phe mutation into the pTR-UFB background 26 using the Quick Change site-Directed Mutagenesis kit (Stratagene). The resulting plasmid was termed pTR-UF6.…”

Section: Construction Of Recombinant Plasmidsmentioning

confidence: 99%

“…Recombinant adeno-associated virus (rAAV) has proven to be a useful vector for efficient and long-term gene transfer in a variety of tissues including lung, 1 muscle, [2][3][4][5] brain, 6 spinal cord, 7 retina 8,9 and liver. 10 rAAV vectors consist of a simple capsid with a single-stranded DNA genome and no viral coding sequences.…”

Section: Introductionmentioning

confidence: 99%

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Recombinant adeno-associated virus purification using novel methods improves infectious titer and yield

et al. 1999

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Conventional methods for rAAV purification that are based vector that is more than 99% pure. More importantly, the on cesium chloride ultracentrifugation have often produced new purification procedures consistently produce rAAV vector preparations of variable quality and resulted in sigstocks with particle-to-infectivity ratios of less than 100, nificant loss of particle infectivity. We report here several which is significantly better than conventional methods. novel purification strategies that involve the use of non-The new protocol increases the overall yield of infectious ionic iodixanol gradients followed by ion exchange or heprAAV by at least 10-fold and allows for the complete purifiarin affinity chromatography by either conventional or cation of rAAV in 1 working day. Several of these methods HPLC columns. These methods result in more than 50%should also be useful for large-scale production. recovery of rAAV from a crude lysate and routinely produce

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Section: Discussionmentioning

confidence: 99%

Section: Figure 2 Titering Of Raav Stocks By the Qc Pcr Assay (A) Or mentioning

confidence: 96%

Section: Construction Of Recombinant Plasmidsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Recombinant adeno-associated virus purification using novel methods improves infectious titer and yield

et al. 1999

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“…The receptor for AAV2 has been characterized and is composed of the heparan sulfate proteoglycan receptor, and at least one co-receptor, FGF-receptor-1 (FGFr1) (Qing et al, 1999;Summerford et al, 1999;Summerford and Samulski, 1998). The relative abundance of these receptors in the brain probably explains why rAAV2 is more efficient in some brain regions such as globus pallidus and substantia nigra than in most other regions Klein et al, 1998). Initially, there was thought to be little immune response to rAAV2 in the brain.…”

Section: Adeno-associated Virusmentioning

confidence: 99%

Viral vectors for in vivo gene transfer in Parkinson's disease: Properties and clinical grade production

Mandel

Bürger

Snyder

2008

Experimental Neurology

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Because Parkinson's disease is a progressive degenerative disorder that is mainly confined to the basal ganglia, gene transfer to deliver therapeutic molecules is an attractive treatment avenue. The present review focuses on direct in vivo gene transfer vectors that have been developed to a degree that they have been successfully used in animal model of Parkinson's disease. Accordingly, the properties of recombinant adenovirus, recombinant adeno-associated virus, herpes simplex virus, and lentivirus are described and contrasted. In order for viral vectors to be developed into clinical grade reagents, they must be manufactured and tested to precise regulatory standards. Indeed, clinical lots of viral vectors can be produced in compliance with current Good Manufacturing Practices (cGMPs) regulations using industry accepted manufacturing methodologies, manufacturing controls, and quality systems. The viral vector properties themselves combined with physiological product formulations facilitate long-term storage and direct in vivo administration.

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Adeno-associated virus-mediated gene delivery

Snyder

1999

J. Gene Med.

102

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Neuron-Specific Transduction in the Rat Septohippocampal or Nigrostriatal Pathway by Recombinant Adeno-associated Virus Vectors

Cited by 229 publications

References 41 publications

Recombinant adeno-associated virus purification using novel methods improves infectious titer and yield

Recombinant adeno-associated virus purification using novel methods improves infectious titer and yield

Viral vectors for in vivo gene transfer in Parkinson's disease: Properties and clinical grade production

Adeno-associated virus-mediated gene delivery

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