Adeno-associated viral vector (AAV)-mediated gene transfer in the red nucleus of the adult rat brain: Comparative analysis of the transduction properties of seven AAV serotypes and lentiviral vectors

Blits, Bas; Derks, Sanne; Twisk, Jaap; Ehlert, Erich; Prins, Jolanda; Verhaagen, Joost

doi:10.1016/j.jneumeth.2009.10.009

Cited by 58 publications

(57 citation statements)

References 56 publications

(77 reference statements)

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“…At the third day after transfection, the medium was changed for lysis buffer (50 mM Tris, 2 mM MgCl 2 , 150 mM NaCl and 0.1% Triton X-100). After DNAse treatment, the crude lysate was loaded onto an iodixanol density gradient (Sigma, St. Louis, MO, USA) and centrifuged for 70 min in a Beckman XL-100K ultracentrifuge at 69 000 rpm at 168C (47). Fractions containing the viral vectors were collected and concentrated using Amicon Ultra-15 concentrators.…”

Section: Aav Gfp and Cre-gfp Vectorsmentioning

confidence: 99%

Targeted ablation of Crb2 in photoreceptor cells induces retinitis pigmentosa

Alves¹,

Pellissier²,

Vos³

et al. 2014

Human Molecular Genetics

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In humans, the Crumbs homolog-1 (CRB1) gene is mutated in autosomal recessive Leber congenital amaurosis and early-onset retinitis pigmentosa. In mammals, the Crumbs family is composed of: CRB1, CRB2, CRB3A and CRB3B. Recently, we showed that removal of mouse Crb2 from retinal progenitor cells, and consequent removal from Mü ller glial and photoreceptor cells, results in severe and progressive retinal degeneration with concomitant loss of retinal function that mimics retinitis pigmentosa due to mutations in the CRB1 gene. Here, we studied the effects of cell-type-specific loss of CRB2 from the developing mouse retina using targeted conditional deletion of Crb2 in photoreceptors or Mü ller cells. We analyzed the consequences of targeted loss of CRB2 in the adult mouse retina using adeno-associated viral vectors encoding Cre recombinase and short hairpin RNA against Crb2. In vivo retinal imaging by means of optical coherence tomography on retinas lacking CRB2 in photoreceptors showed progressive thinning of the photoreceptor layer and cellular mislocalization. Electroretinogram recordings under scotopic conditions showed severe attenuation of the a-wave, confirming the degeneration of photoreceptors. Retinas lacking CRB2 in developing photoreceptors showed early onset of abnormal lamination, whereas retinas lacking CRB2 in developing Mü ller cells showed late onset retinal disorganization. Our data suggest that in the developing retina, CRB2 has redundant functions in Mü ller glial cells, while CRB2 has essential functions in photoreceptors. Our data suggest that short-term loss of CRB2 in adult mouse photoreceptors, but not in Mü ller glial cells, causes sporadic loss of adhesion between photoreceptors and Mü ller cells.

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Section: Aav Gfp and Cre-gfp Vectorsmentioning

confidence: 99%

Targeted ablation of Crb2 in photoreceptor cells induces retinitis pigmentosa

Alves¹,

Pellissier²,

Vos³

et al. 2014

Human Molecular Genetics

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“…Both lentiviral and rAAV vectors have been used to deliver transgenes into the nigrostriatal system for purposes of disease modeling [6][7][8][9][10]24 or gene therapy. 12,25 rAAV vectors are particularly attractive for targeting the substantia nigra (SN) [26][27][28][29][30][31][32] because of their high titers and tropism for neurons, while inducing minimal immune responses in the brain. rAAV vectors are derived from a 4.7-kb singlestranded DNA virus that belongs to the family of the parvoviridae.…”

Section: Introductionmentioning

confidence: 99%

Efficient and stable transduction of dopaminergic neurons in rat substantia nigra by rAAV 2/1, 2/2, 2/5, 2/6.2, 2/7, 2/8 and 2/9

et al. 2011

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Dysfunction of the nigrostriatal system is the major cause of Parkinson's disease (PD). This brain region is therefore an important target for gene delivery aiming at disease modeling and gene therapy. Recombinant adeno-associated viral (rAAV) vectors have been developed as efficient vehicles for gene transfer into the central nervous system. Recently, several serotypes have been described, with varying tropism for brain transduction. In light of the further development of a viral vector-mediated rat model for PD, we performed a comprehensive comparison of the transduction and tropism for dopaminergic neurons (DNs) in the adult Wistar rat substantia nigra (SN) of seven rAAV vector serotypes (rAAV 2/1, 2/2, 2/5, 2/6.2, 2/7, 2/8 and 2/9). All vectors were normalized by titer and volume, and stereotactically injected into the SN. Gene expression was assessed non-invasively and quantitatively in vivo by bioluminescence imaging at 2 and 5 weeks after injection, and was found to be stable over time. Immunohistochemistry at 6 weeks following injection revealed the most widespread enhanced green fluorescence protein expression and the highest number of positive nigral cells using rAAV 2/7, 2/9 and 2/1. The area transduced by rAAV 2/8 was smaller, but nevertheless almost equal numbers of nigral cells were targeted. Detailed confocal analysis revealed that serotype 2/7, 2/9, 2/1 and 2/8 transduced at least 70% of the DNs. In conclusion, these results show that various rAAV serotypes efficiently transduce nigral DNs, but significant differences in transgene expression pattern and level were observed.

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“…This study confirms a more efficient transduction of AAV2/1 compared to AAV2/8 in the VMH. As AAV2/5 is shown to effectively transduce other brain structures [4], [8], [9], this serotype was examined for its transduction efficiency in the VMH. AAV2/5 was found to be equally effective in transducing the VMH as AAV2/1, which corresponds to Burger et al, who compared these serotypes in rat striatum, hippocampus and substantia nigra [4].…”

Section: Discussionmentioning

confidence: 99%

“…Since different serotypes infect different and overlapping types of cells, the AAV2 vector has been pseudotyped in capsids from different AAV serotypes. AAV2/1, AAV2/5 and AAV2/8 have been shown to effectively transduce rat hypothalamus [3], striatum [4], [5], hippocampus [4], [6], [7], substantia nigra [4], [6], [8] and red nucleus [9].…”

Section: Introductionmentioning

confidence: 99%

Recombinant Adeno-Associated Virus: Efficient Transduction of the Rat VMH and Clearance from Blood

et al. 2014

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To promote the efficient and safe application of adeno-associated virus (AAV) vectors as a gene transfer tool in the central nervous system (CNS), transduction efficiency and clearance were studied for serotypes commonly used to transfect distinct areas of the brain. As AAV2 was shown to transduce only small volumes in several brain regions, this study compares the transduction efficiency of three AAV pseudotyped vectors, namely AAV2/1, AAV2/5 and AAV2/8, in the ventromedial nucleus of the hypothalamus (VMH). No difference was found between AAV2/1 and AAV2/5 in transduction efficiency. Both AAV2/1 and AAV2/5 achieved a higher transduction rate than AAV2/8. One hour after virus administration to the brain, no viral particles could be traced in blood, indicating that no or negligible numbers of virions crossed the blood-brain barrier. In order to investigate survival of AAV in blood, clearance was determined following systemic AAV administration. The half-life of AAV2/1, AAV2/2, AAV2/5 and AAV2/8 was calculated by determining virus clearance rates from blood after systemic injection. The half-life of AAV2/2 was 4.2 minutes, which was significantly lower than the half-lives of AAV2/1, AAV2/5 and AAV2/8. With a half-life of more than 11 hours, AAV2/8 particles remained detectable in blood significantly longer than AAV2/5. We conclude that application of AAV in the CNS is relatively safe as no AAV particles are detectable in blood after injection into the brain. With a half-life of 1.67 hours of AAV2/5, a systemic injection with 1×109 genomic copies of AAV would be fully cleared from blood after 2 days.

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Adeno-associated viral vector (AAV)-mediated gene transfer in the red nucleus of the adult rat brain: Comparative analysis of the transduction properties of seven AAV serotypes and lentiviral vectors

Cited by 58 publications

References 56 publications

Targeted ablation of Crb2 in photoreceptor cells induces retinitis pigmentosa

Targeted ablation of Crb2 in photoreceptor cells induces retinitis pigmentosa

Efficient and stable transduction of dopaminergic neurons in rat substantia nigra by rAAV 2/1, 2/2, 2/5, 2/6.2, 2/7, 2/8 and 2/9

Recombinant Adeno-Associated Virus: Efficient Transduction of the Rat VMH and Clearance from Blood

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