Biochemical Investigation of Active Intracellular Transport of Polymeric Gene-Delivery Vectors

2008 ◽  
Vol 97 (4) ◽  
pp. 1399-1413 ◽  
Author(s):  
David M. Drake ◽  
Daniel W. Pack
Keyword(s):  
Gene Delivery ◽  
Intracellular Transport ◽  
Biochemical Investigation ◽  
Gene Delivery Vectors ◽  
Polymeric Gene Delivery

scholarly journals Competitive binding and molecular crowding regulate the cytoplasmic interactome of non-viral polymeric gene delivery vectors

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Aji Alex M. Raynold ◽  
Danyang Li ◽  
Lan Chang ◽  
Julien E. Gautrot
Keyword(s):  
Gene Delivery ◽  
Molecular Mechanisms ◽  
Transfection Efficiency ◽  
Competitive Binding ◽  
Molecular Crowding ◽  
Gene Delivery Vectors ◽  
Polymeric Gene Delivery ◽  
Associated Proteins ◽  
Improved Design

AbstractIn contrast to the processes controlling the complexation, targeting and uptake of polycationic gene delivery vectors, the molecular mechanisms regulating their cytoplasmic dissociation remains poorly understood. Upon cytosolic entry, vectors become exposed to a complex, concentrated mixture of molecules and biomacromolecules. In this report, we characterise the cytoplasmic interactome associated with polycationic vectors based on poly(dimethylaminoethyl methacrylate) (PDMAEMA) and poly(2-methacrylolyloxyethyltrimethylammonium chloride) (PMETAC) brushes. To quantify the contribution of different classes of low molar mass molecules and biomacromolecules to RNA release, we develop a kinetics model based on competitive binding. Our results identify the importance of competition from highly charged biomacromolecules, such as cytosolic RNA, as a primary regulator of RNA release. Importantly, our data indicate the presence of ribosome associated proteins, proteins associated with translation and transcription factors that may underly a broader impact of polycationic vectors on translation. In addition, we bring evidence that molecular crowding modulates competitive binding and demonstrate how the modulation of such interactions, for example via quaternisation or the design of charge-shifting moieties, impacts on the long-term transfection efficiency of polycationic vectors. Understanding the mechanism regulating cytosolic dissociation will enable the improved design of cationic vectors for long term gene release and therapeutic efficacy.

scholarly journals Balancing protection and release of DNA: tools to address a bottleneck of non-viral gene delivery

2009 ◽  
Vol 7 (suppl_1) ◽  
Author(s):  
Christopher L. Grigsby ◽  
Kam W. Leong
Keyword(s):  
Gene Therapy ◽  
Gene Delivery ◽  
Imaging Techniques ◽  
Model Systems ◽  
Viral Gene ◽  
Dna Protection ◽  
Gene Delivery Vectors ◽  
Viral Gene Therapy ◽  
Polymeric Gene Delivery

Engineering polymeric gene-delivery vectors to release an intact DNA payload at the optimal time and subcellular compartment remains a formidable challenge. An ideal vector would provide total protection of complexed DNA from degradation prior to releasing it efficiently near or within the nucleus of a target cell. While optimization of polymer properties, such as molecular weight and charge density, has proved largely inadequate in addressing this challenge, applying polymeric carriers that respond to temperature, light, pH and redox environment to trigger a switch from a tight, protective complex to a more relaxed interaction favouring release at the appropriate time and place has shown promise. Currently, a paucity of gene carriers able to satisfy the contrary requirements of adequate DNA protection and efficient release contributes to the slow progression of non-viral gene therapy towards clinical translation. This review highlights the promising carrier designs that may achieve an optimal balance of DNA protection and release. It also discusses the imaging techniques and three-dimensional in vitro models that can help study these two barriers in the non-viral gene transfer process. Ultimately, efficacious non-viral gene therapy will depend on the combination of intelligent material design, innovative imaging techniques and sophisticated in vitro model systems to facilitate the rational design of polymeric gene-delivery vectors.

In silico prediction of the in vitro behavior of polymeric gene delivery vectors

Nanoscale ◽  
2021 ◽  
Author(s):  
Nina Bono ◽  
Bárbara Coloma Smith ◽  
Francesca Moreschi ◽  
Alberto Redaelli ◽  
Alfonso Gautieri ◽  
...  
Keyword(s):  
Gene Delivery ◽  
In Silico ◽  
Screening Tool ◽  
Test Bench ◽  
In Silico Prediction ◽  
Gene Delivery Vectors ◽  
Polymeric Gene Delivery

The authors describe a novel screening tool to test bench non-viral transfectants enabling to foresee the most suitable conditions for the complexation of relevant siRNA-polycation assemblies.

Recent advances in the analysis full/empty capsid ratio and genome integrity of adeno-associated virus (AAV) gene delivery vectors

2020 ◽  
Vol 20 ◽  
Author(s):  
L. Hajba ◽  
A. Guttman
Keyword(s):  
Gene Delivery ◽  
High Efficiency ◽  
Analytical Techniques ◽  
Viral Gene ◽  
Adeno Associated Virus ◽  
Gene Delivery Vectors ◽  
Empty Capsid ◽  
Purification Methods ◽  
Viral Gene Delivery

: Adeno-associated virus (AAV) is one of the most promising viral gene delivery vectors with long-term gene expression and disease correction featuring high efficiency and excellent safety in human clinical trials. During the production of AAV vectors,there are several quality control (QC)parameters that should be rigorously monitored to comply with clini-cal safety and efficacy. This review gives a short summary of the most frequently used AVV production and purification methods,focusing on the analytical techniques applied to determine the full/empty capsid ratio and the integrity of the encapsidated therapeutic DNA of the products.

pH-sensitive N,N-(dimethyl)-N-alkanamine-N-oxides as gene delivery vectors

2017 ◽  
Vol 71 (9) ◽  
pp. 1739-1748 ◽  
Author(s):  
Gilda Liskayová ◽  
Lukáš Hubčík ◽  
Katarína Šišková ◽  
Ingrid Pauliková ◽  
Dominika Galliková ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Ph Sensitive ◽  
Gene Delivery Vectors

scholarly journals Quantification of cellular and nuclear uptake rates of polymeric gene delivery nanoparticles and DNA plasmids via flow cytometry

2016 ◽  
Vol 37 ◽  
pp. 120-130 ◽  
Author(s):  
Corey J. Bishop ◽  
Rebecca L. Majewski ◽  
Toni-Rose M. Guiriba ◽  
David R. Wilson ◽  
Nupura S. Bhise ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Gene Delivery ◽  
Nuclear Uptake ◽  
Uptake Rates ◽  
Polymeric Gene Delivery

Structure–activity relationship studies of symmetrical cationic bolasomes as non-viral gene vectors

2016 ◽  
Vol 4 (33) ◽  
pp. 5575-5584 ◽  
Author(s):  
Zheng Huang ◽  
Yi-Mei Zhang ◽  
Qian Cheng ◽  
Ji Zhang ◽  
Yan-Hong Liu ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Structure Activity Relationship ◽  
Activity Relationship ◽  
Viral Gene ◽  
Structure Activity ◽  
Gene Delivery Vectors ◽  
Gene Vectors

Bolalipids based on lysine or cyclen headgroups were synthesized and their structure–activity relationship as gene delivery vectors was studied.

Innate immune response induced by gene delivery vectors

2008 ◽  
Vol 354 (1-2) ◽  
pp. 9-15 ◽  
Author(s):  
Haruna Sakurai ◽  
Kenji Kawabata ◽  
Fuminori Sakurai ◽  
Shinsaku Nakagawa ◽  
Hiroyuki Mizuguchi
Keyword(s):  
Immune Response ◽  
Gene Delivery ◽  
Innate Immune Response ◽  
Innate Immune ◽  
Gene Delivery Vectors
Sign in / Sign up

Export Citation Format

Share Document