Synthesis of adenoviral targeting molecules by intein-mediated protein ligation

Nyanguile, Origène; Dancik, Chanteé M.; Blakemore, Judith I.; Mulgrew, Kathy; Kaleko, Michael; Stevenson, Susan C.

doi:10.1038/sj.gt.3301989

Cited by 18 publications

(11 citation statements)

References 45 publications

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“…capsulatus (Bath) Mu-like prophage could be engineered to resemble the Mu derivatives, which have been excellent tools for random mutagenesis in other species (Casadaban and Cohen 1979). Conditional protein splicing via inteins is used as a tool for protein engineering, drug therapy, and vaccine development (Humphries et al 2002; Mootz et al 2003; Nyanguile et al 2003). The putative inteins could be designed as a tool either for generating protein material for vaccination of salmon (or other animals feeding on M.…”

Section: Resultsmentioning

confidence: 99%

Genomic Insights into Methanotrophy: The Complete Genome Sequence of Methylococcus capsulatus (Bath)

Ward¹,

Larsen

Sakwa³

et al. 2004

PLoS Biol

301

278

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Methanotrophs are ubiquitous bacteria that can use the greenhouse gas methane as a sole carbon and energy source for growth, thus playing major roles in global carbon cycles, and in particular, substantially reducing emissions of biologically generated methane to the atmosphere. Despite their importance, and in contrast to organisms that play roles in other major parts of the carbon cycle such as photosynthesis, no genome-level studies have been published on the biology of methanotrophs. We report the first complete genome sequence to our knowledge from an obligate methanotroph, Methylococcus capsulatus (Bath), obtained by the shotgun sequencing approach. Analysis revealed a 3.3-Mb genome highly specialized for a methanotrophic lifestyle, including redundant pathways predicted to be involved in methanotrophy and duplicated genes for essential enzymes such as the methane monooxygenases. We used phylogenomic analysis, gene order information, and comparative analysis with the partially sequenced methylotroph Methylobacterium extorquens to detect genes of unknown function likely to be involved in methanotrophy and methylotrophy. Genome analysis suggests the ability of M. capsulatus to scavenge copper (including a previously unreported nonribosomal peptide synthetase) and to use copper in regulation of methanotrophy, but the exact regulatory mechanisms remain unclear. One of the most surprising outcomes of the project is evidence suggesting the existence of previously unsuspected metabolic flexibility in M. capsulatus, including an ability to grow on sugars, oxidize chemolithotrophic hydrogen and sulfur, and live under reduced oxygen tension, all of which have implications for methanotroph ecology. The availability of the complete genome of M. capsulatus (Bath) deepens our understanding of methanotroph biology and its relationship to global carbon cycles. We have gained evidence for greater metabolic flexibility than was previously known, and for genetic components that may have biotechnological potential.

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Section: Resultsmentioning

confidence: 99%

Genomic Insights into Methanotrophy: The Complete Genome Sequence of Methylococcus capsulatus (Bath)

Ward¹,

Larsen

Sakwa³

et al. 2004

PLoS Biol

301

278

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“…Type 5 replication-deficient adenovirus (Ad) encoding green fluorescent protein (GFP) under the control of CMV promoter ( GFP Ad) was obtained from the Gene Vector Core Facility of the University of Pennsylvania. The human recombinant D1 domain of the Coxsackie-Ad receptor (CAR) was prepared as described elsewhere (12). …”

Section: Methodsmentioning

confidence: 99%

Formulation and In Vitro Characterization of Composite Biodegradable Magnetic Nanoparticles for Magnetically Guided Cell Delivery

et al. 2012

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Purpose Cells modified with magnetically responsive nanoparticles (MNP) can provide the basis for novel targeted therapeutic strategies. However, improvements are required in the MNP design and cell treatment protocols to provide adequate magnetic properties in balance with acceptable cell viability and function. This study focused on select variables controlling the uptake and cell compatibility of biodegradable polymer-based MNP in cultured endothelial cells. Methods Fluorescent-labeled MNP were formed using magnetite and polylactide as structural components. Their magnetically driven sedimentation and uptake were studied fluorimetrically relative to cell viability in comparison to non-magnetic control conditions. The utility of surface-activated MNP forming affinity complexes with replication-deficient adenovirus (Ad) for transduction achieved concomitantly with magnetic cell loading was examined using the green fluorescent protein reporter. Results A high-gradient magnetic field was essential for sedimentation and cell binding of albumin-stabilized MNP, the latter being rate-limiting in the MNP loading process. Cell loading up to 160 pg iron oxide per cell was achievable with cell viability >90%. Magnetically driven uptake of MNP-Ad complexes can provide high levels of transgene expression potentially useful for a combined cell/gene therapy. Conclusions Magnetically responsive endothelial cells for targeted delivery applications can be obtained rapidly and efficiently using composite biodegradable MNP.

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“…When mixed with adenoviral particles, the resulting FA-Fab conjugate (carrying a reporter gene) was shown to redirect adenoviral infection of KB cells via the FR at a level comparable to that achieved by native adenoviral infection. Similarly, Nyanguile et al [100] have used a novel intein-mediated protein ligation (IPL) technique to link FA to CAR D1, a 14 kDa protein that binds to the Ad fiber knob domain. When FA-CAR D1-modified adenovirus was incubated with KB cells, FR-specific transduction of the cells was observed.…”

Section: Adenovirus-based Formulationsmentioning

confidence: 98%

Targeting Therapeutic and Imaging Agents to Folate Receptor Positive Tumors

Reddy

Allagadda²,

Leamon³

2005

CPB

111

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The membrane-bound folate receptor (FR) is overexpressed on a wide range of human cancers, such as those originating in ovary, lung, breast, endometrium, kidney and brain. The vitamin folic acid is a high affinity ligand of the FR which retains its receptor binding properties when conjugated to other molecules. Consequently, "folate targeting" technology has successfully been applied for the delivery of protein toxins, chemotherapeutic agents, radio-imaging and therapeutic agents, MRI contrast agents, liposomes, gene transfer vectors, antisense oligonucleotides, ribozymes and immunotherapeutic agents to FR-positive cancers. These folate-bearing delivery systems have produced major enhancements in cancer cell specificity and selectivity over their non-targeted formulation counterparts. Hence, it is hopeful that this targeting strategy will lead to improvements in the safety and efficacy of clinically-relevant anti-cancer agents. Therefore, the focus of this review will be to highlight the current status of folate-targeted technology with particular emphasis on the recent advances in this field as well as possible directions for future development.

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Synthesis of adenoviral targeting molecules by intein-mediated protein ligation

Cited by 18 publications

References 45 publications

Genomic Insights into Methanotrophy: The Complete Genome Sequence of Methylococcus capsulatus (Bath)

Genomic Insights into Methanotrophy: The Complete Genome Sequence of Methylococcus capsulatus (Bath)

Formulation and In Vitro Characterization of Composite Biodegradable Magnetic Nanoparticles for Magnetically Guided Cell Delivery

Targeting Therapeutic and Imaging Agents to Folate Receptor Positive Tumors

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