Bosch etching for the creation of a 3D nanoelectroporation system for high throughput gene delivery

2015 ◽  
Vol 33 (6) ◽  
pp. 06F903 ◽  
Author(s):  
Paul Bertani ◽  
Wu Lu ◽  
Lingqian Chang ◽  
Daniel Gallego-Perez ◽  
Ly James Lee ◽  
...  
Keyword(s):  
Gene Delivery ◽  
High Throughput ◽  
The Creation

High throughput semi-automated extraction method for the creation of a collection from microbial fermentations of fungi and Actinomycetes

Planta Medica ◽  
2008 ◽  
Vol 74 (09) ◽  
Author(s):  
JR Tormo ◽  
N Tabanera ◽  
D Conway ◽  
P Ramos ◽  
A Redondo ◽  
...  
Keyword(s):  
High Throughput ◽  
Extraction Method ◽  
Automated Extraction ◽  
The Creation

scholarly journals High-Throughput Platform for Efficient Chemical Transfection, Virus Packaging, and Transduction

Micromachines ◽  
2019 ◽  
Vol 10 (6) ◽  
pp. 387
Author(s):  
Jianxiong Zhang ◽  
Yawei Hu ◽  
Xiaoqing Wang ◽  
Peng Liu ◽  
Xiaofang Chen
Keyword(s):  
Gene Delivery ◽  
High Throughput ◽  
Large Scale ◽  
Cell Types ◽  
Long Term Storage ◽  
Efficient Gene ◽  
Genetic Modifications ◽  
Media Change ◽  
Virus Packaging ◽  
Nih 3T3

Intracellular gene delivery is normally required to study gene functions. A versatile platform able to perform both chemical transfection and viral transduction to achieve efficient gene modification in most cell types is needed. Here we demonstrated that high throughput chemical transfection, virus packaging, and transduction can be conducted efficiently on our previously developed superhydrophobic microwell array chip (SMAR-chip). A total of 169 chemical transfections were successfully performed on the chip in physically separated microwells through a few simple steps, contributing to the convenience of DNA delivery and media change on the SMAR-chip. Efficiencies comparable to the traditional transfection in multi-well plates (~65%) were achieved while the manual operations were largely reduced. Two transfection procedures, the dry method amenable for the long term storage of the transfection material and the wet method for higher efficiencies were developed. Multiple transfections in a scheduled manner were performed to further increase the transfection efficiencies or deliver multiple genes at different time points. In addition, high throughput virus packaging integrated with target cell transduction were also proved which resulted in a transgene expression efficiency of >70% in NIH 3T3 cells. In summary, the SMAR-chip based high throughput gene delivery is efficient and versatile, which can be used for large scale genetic modifications in a variety of cell types.

scholarly journals Identification and Validation of Small Molecules That Enhance Recombinant Adeno-associated Virus Transduction following High-Throughput Screens

2016 ◽  
Vol 90 (16) ◽  
pp. 7019-7031 ◽  
Author(s):  
Sarah C. Nicolson ◽  
Chengwen Li ◽  
Matthew L. Hirsch ◽  
Vincent Setola ◽  
R. Jude Samulski
Keyword(s):  
Gene Therapy ◽  
Gene Delivery ◽  
High Throughput ◽  
Adeno Associated Virus ◽  
Virus Gene ◽  
Diploid Cells ◽  
High Throughput Screens ◽  
Group 1

ABSTRACTWhile the recent success of adeno-associated virus (AAV)-mediated gene therapy in clinical trials is promising, challenges still face the widespread applicability of recombinant AAV(rAAV). A major goal is to enhance the transduction efficiency of vectors in order to achieve therapeutic levels of gene expression at a vector dose that is below the immunological response threshold. In an attempt to identify novel compounds that enhance rAAV transduction, we performed two high-throughput screens comprising 2,396 compounds. We identified 13 compounds that were capable of enhancing transduction, of which 12 demonstrated vector-specific effects and 1 could also enhance vector-independent transgene expression. Many of these compounds had similar properties and could be categorized into five groups: epipodophyllotoxins (group 1), inducers of DNA damage (group 2), effectors of epigenetic modification (group 3), anthracyclines (group 4), and proteasome inhibitors (group 5). We optimized dosing for the identified compounds in several immortalized human cell lines as well as normal diploid cells. We found that the group 1 epipodophyllotoxins (teniposide and etoposide) consistently produced the greatest transduction enhancement. We also explored transduction enhancement among single-stranded, self-complementary, and fragment vectors and found that the compounds could impact fragmented rAAV2 transduction to an even greater extent than single-stranded vectors.In vivoanalysis of rAAV2 and all of the clinically relevant compounds revealed that, consistent with ourin vitroresults, teniposide exhibited the greatest level of transduction enhancement. Finally, we explored the capability of teniposide to enhance transduction of fragment vectorsin vivousing an AAV8 capsid that is known to exhibit robust liver tropism. Consistent with ourin vitroresults, teniposide coadministration greatly enhanced fragmented rAAV8 transduction at 48 h and 8 days. This study provides a foundation based on the rAAV small-molecule screen methodology, which is ideally used for more-diverse libraries of compounds that can be tested for potentiating rAAV transduction.IMPORTANCEThis study seeks to enhance the capability of adeno-associated viral vectors for therapeutic gene delivery applicable to the treatment of diverse diseases. To do this, a comprehensive panel of FDA-approved drugs were tested in human cells and in animal models to determine if they increased adeno-associated virus gene delivery. The results demonstrate that particular groups of drugs enhance adeno-associated virus gene delivery by unknown mechanisms. In particular, the enhancement of gene delivery was approximately 50 to 100 times better with than without teniposide, a compound that is also used as chemotherapy for cancer. Collectively, these results highlight the potential for FDA-approved drug enhancement of adeno-associated virus gene therapy, which could result in safe and effective treatments for diverse acquired or genetic diseases.

An encoded viral micropatch for multiplex cell-based assays through localized gene delivery

Lab on a Chip ◽  
2017 ◽  
Vol 17 (14) ◽  
pp. 2435-2442 ◽  
Author(s):  
Sangkwon Han ◽  
Hyung Jong Bae ◽  
Su Deok Kim ◽  
Wook Park ◽  
Sunghoon Kwon
Keyword(s):  
Gene Expression ◽  
Gene Delivery ◽  
High Throughput ◽  
Target Gene ◽  
Target Gene Expression

An encoded viral micropatch is presented for localized target gene expression for high-throughput, high-content cell-based assays.

Parallel High-Throughput Screening of Polymer Vectors for Nonviral Gene Delivery: Evaluation of Structure–Property Relationships of Transfection

2013 ◽  
Vol 15 (9) ◽  
pp. 475-482 ◽  
Author(s):  
Alexandra C. Rinkenauer ◽  
Antje Vollrath ◽  
Anja Schallon ◽  
Lutz Tauhardt ◽  
Kristian Kempe ◽  
...  
Keyword(s):  
Gene Delivery ◽  
High Throughput ◽  
High Throughput Screening ◽  
Nonviral Gene Delivery ◽  
Structure Property ◽  
Structure Property Relationships

Emission enhancement and polarization of semiconductor quantum dots with nanoimprinted plasmonic cavities: towards scalable fabrication of plasmon-exciton displays

Nanoscale ◽  
2015 ◽  
Vol 7 (33) ◽  
pp. 13816-13821 ◽  
Author(s):  
Jasper J. Cadusch ◽  
Evgeniy Panchenko ◽  
Nicholas Kirkwood ◽  
Timothy D. James ◽  
Brant C. Gibson ◽  
...  
Keyword(s):  
Quantum Dots ◽  
High Throughput ◽  
Semiconductor Quantum Dots ◽  
Emission Enhancement ◽  
The Creation ◽  
Nanofabrication Technique ◽  
And Control

An application of a high-throughput nanofabrication technique to the creation of a plasmonic metasurface for the enhancement and control of radiation by quantum dots.

scholarly journals Physical and mechanical cues affecting biomaterial-mediated plasmid DNA delivery: insights into non-viral delivery systems

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Valeria Graceffa
Keyword(s):  
Gene Delivery ◽  
Delivery Systems ◽  
Cellular Adhesion ◽  
Viral Gene ◽  
3D Scaffolds ◽  
Cytoplasmic Transport ◽  
The Creation ◽  
Topographical Cues ◽  
Endocytic Pathways

Abstract Background Whilst traditional strategies to increase transfection efficiency of non-viral systems aimed at modifying the vector or the polyplexes/lipoplexes, biomaterial-mediated gene delivery has recently sparked increased interest. This review aims at discussing biomaterial properties and unravelling underlying mechanisms of action, for biomaterial-mediated gene delivery. DNA internalisation and cytoplasmic transport are initially discussed. DNA immobilisation, encapsulation and surface-mediated gene delivery (SMD), the role of extracellular matrix (ECM) and topographical cues, biomaterial stiffness and mechanical stimulation are finally outlined. Main text Endocytic pathways and mechanisms to escape the lysosomal network are highly variable. They depend on cell and DNA complex types but can be diverted using appropriate biomaterials. 3D scaffolds are generally fabricated via DNA immobilisation or encapsulation. Degradation rate and interaction with the vector affect temporal patterns of DNA release and transgene expression. In SMD, DNA is instead coated on 2D surfaces. SMD allows the incorporation of topographical cues, which, by inducing cytoskeletal re-arrangements, modulate DNA endocytosis. Incorporation of ECM mimetics allows cell type-specific transfection, whereas in spite of discordances in terms of optimal loading regimens, it is recognised that mechanical loading facilitates gene transfection. Finally, stiffer 2D substrates enhance DNA internalisation, whereas in 3D scaffolds, the role of stiffness is still dubious. Conclusion Although it is recognised that biomaterials allow the creation of tailored non-viral gene delivery systems, there still are many outstanding questions. A better characterisation of endocytic pathways would allow the diversion of cell adhesion processes and cytoskeletal dynamics, in order to increase cellular transfection. Further research on optimal biomaterial mechanical properties, cell ligand density and loading regimens is limited by the fact that such parameters influence a plethora of other different processes (e.g. cellular adhesion, spreading, migration, infiltration, and proliferation, DNA diffusion and release) which may in turn modulate gene delivery. Only a better understanding of these processes may allow the creation of novel robust engineered systems, potentially opening up a whole new area of biomaterial-guided gene delivery for non-viral systems.

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