Single Cell Kinetics of Intracellular, Nonviral, Nucleic Acid Delivery Vehicle Acidification and Trafficking

2005 ◽  
Vol 16 (4) ◽  
pp. 986-994 ◽  
Author(s):  
Rajan P. Kulkarni ◽  
Swaroop Mishra ◽  
Scott E. Fraser ◽  
Mark E. Davis
Keyword(s):  
Nucleic Acid ◽  
Single Cell ◽  
Cell Kinetics ◽  
Nucleic Acid Delivery ◽  
Delivery Vehicle ◽  
Kinetics Of

scholarly journals Single Cell Kinetics of Phenotypic Switching in the Arabinose Utilization System of E. coli

PLoS ONE ◽  
2014 ◽  
Vol 9 (2) ◽  
pp. e89532 ◽  
Author(s):  
Georg Fritz ◽  
Judith A. Megerle ◽  
Sonja A. Westermayer ◽  
Delia Brick ◽  
Ralf Heermann ◽  
...  
Keyword(s):  
Single Cell ◽  
Cell Kinetics ◽  
Phenotypic Switching ◽  
E Coli ◽  
Kinetics Of

Single-cell kinetics of a repressilator when implemented in a single-copy plasmid

2015 ◽  
Vol 11 (7) ◽  
pp. 1939-1945 ◽  
Author(s):  
Samuel M. D. Oliveira ◽  
Jerome G. Chandraseelan ◽  
Antti Häkkinen ◽  
Nadia S. M. Goncalves ◽  
Olli Yli-Harja ◽  
...  
Keyword(s):  
Single Cell ◽  
Cell Kinetics ◽  
Single Copy ◽  
F Plasmid ◽  
Kinetics Of

We constructed a single-copy repressilator (SCR) by implementing the original repressilator circuit on a single-copy F-plasmid.

scholarly journals Correlation of mRNA delivery timing and protein expression in lipid-based transfection

2019 ◽  
Author(s):  
A. Reiser ◽  
D. Woschée ◽  
N. Mehrotra ◽  
R. Krzysztoń ◽  
H. H. Strey ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Protein Expression ◽  
Single Cell ◽  
Serum Protein ◽  
Transfection Efficiency ◽  
Viral Gene ◽  
Nucleic Acid Delivery ◽  
Delivery Times ◽  
Time Courses ◽  
Endosomal Release

AbstractNon-viral gene delivery is constrained by the dwell time that most synthetic nucleic acid nanocarriers spend inside endosomal compartments. In order to overcome this endosomal-release bottleneck, methods are required that measure nanocarrier uptake kinetics and transfection efficiency simultaneously. Here, we employ live-cell imaging on single-cell arrays (LISCA) to study the delivery-time distribution of lipid-based mRNA complexes under varied serum conditions. By fitting a translation-maturation model to hundreds of individual eGFP reporter fluorescence time courses, the protein expression onset times and the expression rates after transfection are determined. Using this approach, we find that delivery timing and protein expression rates are not intrinsically correlated at the single-cell level, even though population-averaged values of both parameters conjointly change as a function of increasing external serum protein fraction. Lipofectamine mediated delivery showed decreased transfection efficiency and longer delivery times with increasing serum protein concentration. This is in contrast to ionizable lipid nanoparticles (LNPs) mediated transfer, which showed increased efficiency and faster uptake in the presence of serum. In conclusion, the interdependences of single-cell expression rates and onset timing provide additional clues on uptake and release mechanisms, which are useful for improving nucleic acid delivery.

scholarly journals Single-cell kinetics of siRNA-mediated mRNA degradation

2019 ◽  
Vol 21 ◽  
pp. 102077 ◽  
Author(s):  
Rafał Krzysztoń ◽  
Daniel Woschée ◽  
Anita Reiser ◽  
Gerlinde Schwake ◽  
Helmut H. Strey ◽  
...  
Keyword(s):  
Single Cell ◽  
Cell Kinetics ◽  
Mrna Degradation ◽  
Kinetics Of

scholarly journals Correlation of mRNA delivery timing and protein expression in lipid-based transfection

2019 ◽  
Vol 11 (9) ◽  
pp. 362-371 ◽  
Author(s):  
A Reiser ◽  
D Woschée ◽  
N Mehrotra ◽  
R Krzysztoń ◽  
H H Strey ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Protein Expression ◽  
Single Cell ◽  
Serum Protein ◽  
Transfection Efficiency ◽  
Viral Gene ◽  
Nucleic Acid Delivery ◽  
Delivery Times ◽  
Time Courses ◽  
Endosomal Release

Abstract Non-viral gene delivery is constrained by the dwell time that most synthetic nucleic acid nanocarriers spend inside endosomal compartments. In order to overcome this endosomal-release bottleneck, methods are required that measure nanocarrier uptake kinetics and transfection efficiency simultaneously. Here, we employ live-cell imaging on single-cell arrays (LISCA) to study the delivery-time distribution of lipid-based mRNA complexes under varied serum conditions. By fitting a translation-maturation model to hundreds of individual eGFP reporter fluorescence time courses, the protein expression onset times and the expression rates after transfection are determined. Using this approach, we find that delivery timing and protein expression rates are not intrinsically correlated at the single-cell level, even though population-averaged values of both parameters conjointly change as a function of increasing external serum protein fraction. Lipofectamine-mediated delivery showed decreased transfection efficiency and longer delivery times with increasing serum protein concentration. This is in contrast to ionizable lipid nanoparticle (i-LNP)-mediated transfer, which showed increased efficiency and faster uptake in the presence of serum. In conclusion, the interdependences of single-cell expression rates and onset timing provide additional clues on uptake and release mechanisms, which are useful for improving nucleic acid delivery.

scholarly journals HDL as a drug and nucleic acid delivery vehicle

2015 ◽  
Vol 6 ◽  
Author(s):  
Andras G. Lacko ◽  
Nirupama A. Sabnis ◽  
Bhavani Nagarajan ◽  
Walter J. McConathy
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acid Delivery ◽  
Delivery Vehicle

Non-Viral Nucleic Acid Delivery: Key Challenges and Future Directions

2011 ◽  
Vol 8 (3) ◽  
pp. 235-244 ◽  
Author(s):  
Mahmoud Elsabahy ◽  
Adil Nazarali ◽  
Marianna Foldvari
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acid Delivery ◽  
Viral Nucleic Acid ◽  
Future Directions

Charge‐Conversion Strategies for Nucleic Acid Delivery

2021 ◽  
pp. 2011103
Author(s):  
Kingshuk Dutta ◽  
Ritam Das ◽  
Jewel Medeiros ◽  
Pintu Kanjilal ◽  
S. Thayumanavan
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acid Delivery

scholarly journals Synthetic Histidine-Rich Peptides Inhibit Candida Species and Other Fungi In Vitro: Role of Endocytosis and Treatment Implications

2006 ◽  
Vol 50 (8) ◽  
pp. 2797-2805 ◽  
Author(s):  
Jingsong Zhu ◽  
Paul W. Luther ◽  
Qixin Leng ◽  
A. James Mixson
Keyword(s):  
Nucleic Acid ◽  
Antifungal Activity ◽  
Antifungal Agents ◽  
Fungal Growth ◽  
Nucleic Acid Delivery ◽  
Histatin 5 ◽  
Ph Buffering ◽  
Endosomal Acidification

ABSTRACT A family of histidine-rich peptides, histatins, is secreted by the parotid gland in mammals and exhibits marked inhibitory activity against a number of Candida species. We were particularly interested in the mechanism by which histidine-rich peptides inhibit fungal growth, because our laboratory has synthesized a variety of such peptides for drug and nucleic acid delivery. In contrast to naturally occurring peptides that are linear, peptides made on synthesizers can be varied with respect to their degrees of branching. Using this technology, we explored whether histidine-lysine (HK) polymers of different complexities and degrees of branching affect the growth of several species of Candida. Polymers with higher degrees of branching were progressively more effective against Candida albicans, with the four-branched polymer, H2K4b, most effective. Furthermore, H2K4b accumulated efficiently in C. albicans, which may indicate its ability to transport other antifungal agents intracellularly. Although H2K4b had greater antifungal activity than histatin 5, their mechanisms were similar. Toxicity in C. albicans induced by histatin 5 or branched HK peptides was markedly reduced by 4,4′-diisothiocyanato-stilbene-2,2′-disulfonate, an inhibitor of anion channels. We also determined that bafilomycin A1, an inhibitor of endosomal acidification, significantly decreased the antifungal activity of H2K4b. This suggests that the pH-buffering and subsequent endosomal-disrupting properties of histidine-rich peptides have a role in their antifungal activity. Moreover, the ability of the histidine component of these peptides to disrupt endosomes, which allows their escape from the lysosomal pathway, may explain why these peptides are both effective antifungal agents and nucleic acid delivery carriers.

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