Microfluidic Cell Stretching for Highly Effective Gene Delivery into Hard-to-Transfect Primary Cells

ACS Nano ◽  
2020 ◽  
Vol 14 (11) ◽  
pp. 15094-15106 ◽  
Author(s):  
Jeongsoo Hur ◽  
Inae Park ◽  
Kyung Min Lim ◽  
Junsang Doh ◽  
Ssang-Goo Cho ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Primary Cells ◽  
Cell Stretching ◽  
Highly Effective

Synthesis of Poly(β-amino ester)s Optimized for Highly Effective Gene Delivery

2003 ◽  
Vol 14 (5) ◽  
pp. 979-988 ◽  
Author(s):  
Akin Akinc ◽  
Daniel G. Anderson ◽  
David M. Lynn ◽  
Robert Langer
Keyword(s):  
Gene Delivery ◽  
Amino Ester ◽  
Highly Effective

A highly effective gene delivery vector – hyperbranched poly(2-(dimethylamino)ethyl methacrylate) from in situ deactivation enhanced ATRP

2010 ◽  
Vol 46 (26) ◽  
pp. 4698 ◽  
Author(s):  
Ben Newland ◽  
Hongyun Tai ◽  
Yu Zheng ◽  
Diego Velasco ◽  
Andrea Di Luca ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Gene Delivery Vector ◽  
Ethyl Methacrylate ◽  
Delivery Vector ◽  
Hyperbranched Poly ◽  
Highly Effective

scholarly journals A guide in lentiviral vector production for hard-to-transfect cells, using cardiac-derived c-kit expressing cells as a model system

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
V. Kalidasan ◽  
Wai Hoe Ng ◽  
Oluwaseun Ayodeji Ishola ◽  
Nithya Ravichantar ◽  
Jun Jie Tan ◽  
...  
Keyword(s):  
Stem Cells ◽  
Gene Therapy ◽  
Gene Delivery ◽  
Delivery System ◽  
Lentiviral Vector ◽  
Lentiviral Vectors ◽  
Model System ◽  
Selection Marker ◽  
Target Cells ◽  
Primary Cells

AbstractGene therapy revolves around modifying genetic makeup by inserting foreign nucleic acids into targeted cells via gene delivery methods to treat a particular disease. While the genes targeted play a key role in gene therapy, the gene delivery system used is also of utmost importance as it determines the success of gene therapy. As primary cells and stem cells are often the target cells for gene therapy in clinical trials, the delivery system would need to be robust, and viral-based entries such as lentiviral vectors work best at transporting the transgene into the cells. However, even within lentiviral vectors, several parameters can affect the functionality of the delivery system. Using cardiac-derived c-kit expressing cells (CCs) as a model system, this study aims to optimize lentiviral production by investigating various experimental factors such as the generation of the lentiviral system, concentration method, and type of selection marker. Our findings showed that the 2nd generation system with pCMV-dR8.2 dvpr as the packaging plasmid produced a 7.3-fold higher yield of lentiviral production compared to psPAX2. Concentrating the virus with ultracentrifuge produced a higher viral titer at greater than 5 × 105 infectious unit values/ml (IFU/ml). And lastly, the minimum inhibitory concentration (MIC) of puromycin selection marker was 10 μg/mL and 7 μg/mL for HEK293T and CCs, demonstrating the suitability of antibiotic selection for all cell types. This encouraging data can be extrapolated and applied to other difficult-to-transfect cells, such as different types of stem cells or primary cells.

scholarly journals Highly Effective Gene Transfection In Vivo by Alkylated Polyethylenimine

2011 ◽  
Vol 2011 ◽  
pp. 1-6 ◽  
Author(s):  
Jennifer A. Fortune ◽  
Tatiana I. Novobrantseva ◽  
Alexander M. Klibanov
Keyword(s):  
Gene Expression ◽  
Gene Delivery ◽  
Cellular Uptake ◽  
Alkyl Chain ◽  
Transfection Efficiency ◽  
Gene Transfection ◽  
Mouse Lung ◽  
Highly Effective ◽  
Chain Lengths

We mechanistically explored the effect of increased hydrophobicity of the polycation on the efficacy and specificity of gene delivery in mice. N-Alkylated linear PEIs with varying alkyl chain lengths and extent of substitution were synthesized and characterized by biophysical methods. Their in vivo transfection efficiency, specificity, and biodistribution were investigated. N-Ethylation improves the in vivo efficacy of gene expression in the mouse lung 26-fold relative to the parent polycation and more than quadruples the ratio of expression in the lung to that in all other organs. N-Propyl-PEI was the best performer in the liver and heart (581- and 3.5-fold enhancements, resp.) while N-octyl-PEI improved expression in the kidneys over the parent polymer 221-fold. As these enhancements in gene expression occur without changing the plasmid biodistribution, alkylation does not alter the cellular uptake but rather enhances transfection subsequent to cellular uptake.

Highly effective and slow-biodegradable network-type cationic gene delivery polymer: Small library-like approach synthesis and characterization

Biomaterials ◽  
2006 ◽  
Vol 27 (10) ◽  
pp. 2292-2301 ◽  
Author(s):  
Hyun Jin Kim ◽  
Min Sung Kwon ◽  
Joon Sig Choi ◽  
Seung-Min Yang ◽  
Jae Keun Yoon ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Synthesis And Characterization ◽  
Highly Effective ◽  
Network Type

scholarly journals Transduction of Renal Cells in Vitro and in Vivo by Adeno-Associated Virus Gene Therapy Vectors

1999 ◽  
Vol 10 (9) ◽  
pp. 1908-1915 ◽  
Author(s):  
MICHAEL S. LIPKOWITZ ◽  
BASIL HANSS ◽  
NATALIE TULCHIN ◽  
PATRICIA D. WILSON ◽  
JESSICA C. LANGER ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Therapy ◽  
Gene Delivery ◽  
Mesangial Cells ◽  
Renal Diseases ◽  
Primary Cells ◽  
Renal Cells ◽  
Adeno Associated Virus

Abstract. There has been an increasing interest recently in the possibility of treating renal diseases using gene therapy. The ability to pursue gene therapy for renal diseases has been limited by the availability of an adequate system for gene delivery to the kidney. Adeno-associated virus (AAV) is a defective virus of the parvovirus family that has a number of properties attractive for renal gene delivery: recombinant AAV contains no viral genes; expression of genes delivered by these vectors does not activate cell-mediated immunity; the virus is able to transduce nondividing as well as dividing cells; and both wild-type and recombinant AAV integrate into the host chromosome resulting in long-term gene expression. Studies were performed to determine whether AAV can deliver reporter genes to kidney cells in vitro and in vivo. These studies show that AAV can deliver reporter genes with approximately equal efficiency to human mesangial, proximal tubule, thick ascending limb, collecting tubule, and renal cell carcinoma cells in primary culture. Immortalized mouse mesangial cells are transduced at a much greater efficiency. Transduction can be enhanced by pharmaceutical agents up to sevenfold in primary cells (transducing up to 20% of primary cells per well) and as much as 400-fold in immortalized mesangial cells. AAV delivered in vivo by intraparenchymal injection results in at least 3 mo of reporter gene expression in tubular epithelial, but not glomerular or vascular, cells at the injection site. These data indicate that AAV can deliver genes to renal cells both in vitro and in vivo resulting in prolonged gene expression, and thus AAV can be a useful tool for renal gene delivery.

Application of pulsed-magnetic field enhances non-viral gene delivery in primary cells from different origins

2008 ◽  
Vol 320 (8) ◽  
pp. 1517-1527 ◽  
Author(s):  
Sarah W. Kamau Chapman ◽  
Paul O. Hassa ◽  
Sabine Koch-Schneidemann ◽  
Brigitte von Rechenberg ◽  
Margarethe Hofmann-Amtenbrink ◽  
...  
Keyword(s):  
Magnetic Field ◽  
Gene Delivery ◽  
Pulsed Magnetic Field ◽  
Viral Gene ◽  
Primary Cells ◽  
Viral Gene Delivery ◽  
Different Origins

Electrostatic Ligand Coatings of Nanoparticles Enable Ligand-Specific Gene Delivery to Human Primary Cells

Nano Letters ◽  
2007 ◽  
Vol 7 (4) ◽  
pp. 874-879 ◽  
Author(s):  
Jordan J. Green ◽  
Eugene Chiu ◽  
Elizaveta S. Leshchiner ◽  
Julie Shi ◽  
Robert Langer ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Specific Gene ◽  
Primary Cells ◽  
Human Primary Cells
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