Use of adeno-associated viral vector for delivery of small interfering RNA

Tomar, Raghuvir Singh; Matta, Hittu; Chaudhary, Preet M.

doi:10.1038/sj.onc.1206733

Cited by 181 publications

(92 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Retroviral, 50 adenoviral, 51 adeno-associated viral 52 and lentiviral 40 vectors engineered to express siRNA have been proposed, taking advantage of high rate transfection in vivo into tumor cells. 36,39 Moreover, viral vectors may enable relatively persistent RNAi effect in comparison with transient gene silencing obtained by synthetic siRNA duplexes.…”

Section: Discussionmentioning

confidence: 99%

Therapeutic RNA interference of malignant melanoma by electrotransfer of small interfering RNA targeting Mitf

et al. 2006

View full text Add to dashboard Cite

Microphthalmia-associated transcription factor (Mitf) is critically involved in melanin synthesis as well as differentiation of cells of the melanocytic lineage. Some earlier studies suggested that Mitf is also essential in the survival of melanoma cells, but this notion remains controversial. We synthesized short interfering RNA (siRNA) duplexes corresponding to the mitf sequence and transfected them into B16 melanoma. Lipid-mediated transfection in vitro of Mitfspecific siRNA resulted in specific downregulation of Mitf and of the tyrosinase that is a transcriptional target of Mitf. This treatment also remarkably reduced the viability of melanoma cells by inducing apoptosis. To examine the potential feasibility of RNAi therapy against melanoma, B16 cells were subcutaneously injected into syngenic mice and siRNA was transfected into the pre-established tumor by means of electroporation. The Mitf-specific siRNA drastically reduced outgrowth of subcutaneous melanoma, while nonspecific siRNA failed to affect tumor progression. Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling-based analysis of tumor specimens demonstrated that the tumor cells transfected with Mitf-siRNA effectively underwent apoptosis in vivo. The present results indicate that Mitf plays important roles in melanoma survival. Intratumor electrotransfer of Mitf-specific siRNA may provide a powerful strategy for therapeutic intervention of malignant melanoma.

show abstract

Section: Discussionmentioning

confidence: 99%

Therapeutic RNA interference of malignant melanoma by electrotransfer of small interfering RNA targeting Mitf

et al. 2006

View full text Add to dashboard Cite

show abstract

“…To generate pSUPER-Casp2/ EGFP, the pSUPER/EGFP was digested with BglII and HindIII, and the annealed targeting oligonucleotides ACAGCTGTTGTTGAGCGAA (14) were ligated into the vector. To generate pSUPER-Casp3/EGFP and pSUPER-Casp8/EGFP, the pSUPER/EGFP was digested with BglII and HindIII, and the annealed targeting oligonucleotides TGA-CATCTCGGTCTGGTAC (29) and GTTCCTGAGCCTGGACTAC (30) were ligated into the vector, respectively. The pSUPER-Casp2 (-3 or -8)/EGFP/Neo was generated by inserting the Neo r gene from the pIRESneo2 (Clontech) into the pSUPER-Casp2 (-3 or -8)/EGFP vector.…”

Section: Methodsmentioning

confidence: 99%

Sequential Caspase-2 and Caspase-8 Activation Upstream of Mitochondria during Ceramideand Etoposide-induced Apoptosis

Lin¹,

Chen

Chang³

et al. 2004

Journal of Biological Chemistry

123

113

View full text Add to dashboard Cite

Recently, caspase-2 was shown to act upstream of mitochondria in stress-induced apoptosis. Activation of caspase-8, a key event in death receptor-mediated apoptosis, also has been demonstrated in death receptor-independent apoptosis. The regulation of these initiator caspases, which trigger the mitochondrial apoptotic pathway, is unclear. Here we report a potential regulatory role of caspase-2 on caspase-8 during ceramide-induced apoptosis. Our results demonstrate the sequential events of initiator caspase-2 and caspase-8 activation, Bid cleavage and translocation, and mitochondrial damage followed by downstream caspase-9 and -3 activation and cell apoptosis after ceramide induction in T cell lines. The expression of truncated Bid (tBid) and the reduction in mitochondrial transmembrane potential were blocked by caspase-2 or caspase-8, but not caspase-3, knockdown using an RNA interference technique. Ceramide-induced caspase-8 activation, mitochondrial damage, and apoptosis were blocked in caspase-2 short interfering RNA-expressing cells. Therefore, caspase-2 acts upstream of caspase-8 during ceramide-induced mitochondrial apoptosis. Similarly, sequential caspase-2 and caspase-8 activation upstream of mitochondria was also observed in etoposide-induced apoptosis. These data suggest sequential initiator caspase-2 and caspase-8 activation in the mitochondrial apoptotic pathway induced by ceramide or etoposide.A mitochondrial apoptotic pathway is required for various death stimuli (1, 2). The involvement of initiator caspases before mitochondrial damage has been shown (3, 4). Caspase-8 activation followed by the cleavage of the Bcl-2 family protein Bid, a BH3 domain-containing proapoptotic protein, induces mitochondrial damage during death receptor-mediated apoptosis (5-8). Activation of caspase-8 also has been demonstrated in death receptor-independent apoptosis (9, 10). Caspase-2 recently was shown to act upstream of mitochondria to promote cytochrome c release and apoptosis (11)(12)(13)(14). The regulation of these initiator caspases leading to the mitochondria-mediated apoptotic pathway remains unresolved.Ceramide, a product of sphingolipid metabolism, regulates diverse cellular responses, including proliferation, differentiation, and apoptosis. Although the role of ceramide as a signaling molecule remains debatable (15, 16), some early work suggested that various extracellular agents and stress stimuli, such as tumor necrosis factor-␣ (TNF␣), Fas, chemotherapeutic agents, irradiation, and heat, might cause accumulation of ceramide. Ceramide modulates the biochemical and cellular processes that lead to apoptosis (17-21). The mechanisms by which ceramide regulates apoptotic events have not been fully defined. Ceramide activates a number of enzymes involved in stress-signaling pathways, including both protein kinases and protein phosphatases. A recent study indicates that ceramide-induced neuronal apoptosis is associated with dephosphorylation of Akt, Bcl2-associated death promoter (BAD), forkhead transcriptional ...

show abstract

“…One important feature of AAV2 is that this vector remains confined to the injection site and specifically infects neurons rather than glia (Bartlett et al 1998, Tomar et al 2003, Grimm et al 2005. By inserting an additional expression cassette to encode EGFP for fluorescent detection, we were able to visualize the injection site and AAV transduction in the brain.…”

Section: Discussionmentioning

confidence: 99%

Adeno-associated virus-mediated knockdown of melanocortin-4 receptor in the paraventricular nucleus of the hypothalamus promotes high-fat diet-induced hyperphagia and obesity

Garza

Kim

Liu

et al. 2008

Journal of Endocrinology

View full text Add to dashboard Cite

Pharmacological and genetic studies have suggested that melanocortin-4 receptor (MC4R) signaling in the paraventricular nucleus of hypothalamus (PVN) regulates appetite and energy balance. However, the specific role of MC4R signaling in PVN neurons in these processes remains to be further elucidated in normally developed animals. In the present study, we employed RNA interference to determine whether MC4R knockdown in the PVN modulates food intake and body weight in adult rats. Adeno-associated viral (AAV) vectors encoding short hairpin RNAs targeting MC4R (AAV-shRNA-MC4R) were generated to induce MC4R knockdown in the PVN. By in situ hybridization, we detected a high-level expression of Dicer, a key enzyme required for shRNA-mediated gene silencing, along the entire rostrocaudal extent of the PVN. Bilateral injection of AAV-shRNA-MC4R vectors into the PVN of the adult rat resulted in significant and specific reduction of MC4R mRNA expression. Animals with MC4R knockdown exhibited an increase in food intake and excessive body weight gain when exposed to a high-fat diet. Our results provide evidence that AAV-mediated silencing of MC4R on PVN neurons promotes hyperphagia and obesity in response to the dietary challenge in the adult animal.

show abstract

Use of adeno-associated viral vector for delivery of small interfering RNA

Cited by 181 publications

References 19 publications

Therapeutic RNA interference of malignant melanoma by electrotransfer of small interfering RNA targeting Mitf

Therapeutic RNA interference of malignant melanoma by electrotransfer of small interfering RNA targeting Mitf

Sequential Caspase-2 and Caspase-8 Activation Upstream of Mitochondria during Ceramideand Etoposide-induced Apoptosis

Adeno-associated virus-mediated knockdown of melanocortin-4 receptor in the paraventricular nucleus of the hypothalamus promotes high-fat diet-induced hyperphagia and obesity

Contact Info

Product

Resources

About