scholarly journals DNA hydrogel microspheres and their potential applications for protein delivery and live cell monitoring

2016 ◽  
Vol 10 (3) ◽  
pp. 034112 ◽  
Author(s):  
Taeyoung Kim ◽  
Seongmin Park ◽  
Minhyuk Lee ◽  
Solhee Baek ◽  
Jong Bum Lee ◽  
...  
Keyword(s):  
Protein Delivery ◽  
Live Cell ◽  
Potential Applications ◽  
Cell Monitoring ◽  
Dna Hydrogel ◽  
Hydrogel Microspheres

DNA Hydrogel with Aptamer-Toehold-Based Recognition, Cloaking, and Decloaking of Circulating Tumor Cells for Live Cell Analysis

Nano Letters ◽  
2017 ◽  
Vol 17 (9) ◽  
pp. 5193-5198 ◽  
Author(s):  
Ping Song ◽  
Dekai Ye ◽  
Xiaolei Zuo ◽  
Jiang Li ◽  
Jianbang Wang ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Live Cell ◽  
Cell Analysis ◽  
Dna Hydrogel ◽  
Live Cell Analysis

scholarly journals Live Cell Monitoring of μ-Opioid Receptor-mediated G-protein Activation Reveals Strong Biological Activity of Close Morphine Biosynthetic Precursors

2007 ◽  
Vol 282 (37) ◽  
pp. 27126-27132 ◽  
Author(s):  
Viacheslav O. Nikolaev ◽  
Chotima Boettcher ◽  
Christian Dees ◽  
Moritz Bünemann ◽  
Martin J. Lohse ◽  
...  
Keyword(s):  
Biological Activity ◽  
G Protein ◽  
Opioid Receptor ◽  
Live Cell ◽  
Protein Activation ◽  
G Protein Activation ◽  
Cell Monitoring ◽  
Μ Opioid Receptor

scholarly journals Live Cell Monitoring of hiPSC Generation and Differentiation Using Differential Expression of Endogenous microRNAs

PLoS ONE ◽  
2010 ◽  
Vol 5 (7) ◽  
pp. e11834 ◽  
Author(s):  
Masakazu Kamata ◽  
Min Liang ◽  
Shirley Liu ◽  
Yoshiko Nagaoka ◽  
Irvin S. Y. Chen
Keyword(s):  
Differential Expression ◽  
Live Cell ◽  
Cell Monitoring

scholarly journals Genetically encoded live cell sensor for tyrosinated microtubules

2020 ◽  
Author(s):  
Shubham Kesarwani ◽  
Prakash Lama ◽  
Anchal Chandra ◽  
P. Purushotam Reddy ◽  
AS Jijumon ◽  
...  
Keyword(s):  
Living Cells ◽  
Live Cell ◽  
Live Cells ◽  
Yeast Display ◽  
Post Translational Modification ◽  
Alpha Tubulin ◽  
Microtubule Stability ◽  
Specific Manner ◽  
Potential Applications ◽  
Stability Dynamics

AbstractMicrotubule cytoskeleton exists in various biochemical forms in different cells due to tubulin post-translational modification (PTMs). These PTMs are known to affect microtubule stability, dynamics and interaction with MAPs and motors in a specific manner, widely known as tubulin code hypothesis. At present there exist no tool that can specifically mark tubulin PTMs in live cells, thus severely limiting our understanding of tubulin PTMs. Using yeast display library, we identified a binder against terminal tyrosine of alpha tubulin, a unique PTM site. Extensive characterization validates the robustness and non-perturbing nature of our binder as tyrosination sensor, a live cell tubulin nanobody specific towards tyrosinated or unmodified microtubules. Using which, in real time we followed nocodazole, colchicine and vincristine induced depolymerization events of unmodified microtubules, and found each distinctly perturb microtubule polymer. Together, our work describes the tyrosination sensor and potential applications to study microtubule and PTM processes in living cells.

scholarly journals Engineered type six secretion systems deliver active exogenous effectors and Cre recombinase

2021 ◽  
Author(s):  
Steven Jeremy Hersch ◽  
Linh Lam ◽  
Tao Dong
Keyword(s):  
Protein Delivery ◽  
Cre Recombinase ◽  
Future Application ◽  
Secretion Systems ◽  
Chaperone Protein ◽  
Potential Applications ◽  
Cre Recombination ◽  
Bacterial Type ◽  
Target Effects ◽  
Efficient Transfer

Genetic editing has revolutionized biotechnology but delivery of endonuclease genes as DNA can lead to aberrant integration or overexpression, leading to off-target effects. Here we develop a mechanism to deliver Cre recombinase as a protein by engineering the bacterial type six secretion system (T6SS). Using multiple T6SS fusion proteins, Aeromonas dhakensis or attenuated Vibrio cholerae donor strains, and a gain-of-function cassette for detecting Cre recombination, we demonstrate successful delivery of active Cre directly into recipient cells. Most efficient transfer was achieved using a truncated version of PAAR2 from V. cholerae, resulting in a relatively small (118 amino acid) 'delivery tag'. We further demonstrate the versatility of this system by delivering an exogenous effector, TseC, enabling V. cholerae to kill Pseudomonas aeruginosa. This implicates that P. aeruginosa is naturally resistant to all native effectors of V. cholerae and that the TseC chaperone protein is not required for its activity. Moreover, it demonstrates that the engineered system can improve T6SS efficacy against specific pathogens, proposing future application in microbiome manipulation or as a next-generation antimicrobial. Inexpensive and easy to produce, this protein delivery system has many potential applications ranging from studying T6SS effectors to genetic editing.

Hydrogel microspheres evading alveolar macrophages for sustained pulmonary protein delivery

2019 ◽  
Vol 566 ◽  
pp. 652-661 ◽  
Author(s):  
Moritz Graf ◽  
Christian E. Ziegler ◽  
Manuel Gregoritza ◽  
Achim M. Goepferich
Keyword(s):  
Alveolar Macrophages ◽  
Protein Delivery ◽  
Hydrogel Microspheres

Design and Characterization of a Fluorescent Reporter Enabling Live-cell Monitoring of MCT8 Expression

2021 ◽  
Author(s):  
Adina Sophie Graffunder ◽  
Sarah Paisdzior ◽  
Robert Opitz ◽  
Kostja Renko ◽  
Peter Kühnen ◽  
...  
Keyword(s):  
Expression Patterns ◽  
Functional Characterization ◽  
Live Cell ◽  
Monocarboxylate Transporter ◽  
Start Codon ◽  
Future Application ◽  
Gene Assay ◽  
Cell Monitoring

AbstractThe monocarboxylate transporter 8 (MCT8) is a specific thyroid hormone transporter and plays an essential role in fetal development. Inactivating mutations in the MCT8 encoding gene SLC16A2 (solute carrier family 16, member 2) lead to the Allan-Herndon-Dudley syndrome, a condition presenting with severe endocrinological and neurological phenotypes. However, the cellular distribution pattern and dynamic expression profile are still not well known for early human neural development. Objective Development and characterization of fluorescent MCT8 reporters that would permit live-cell monitoring of MCT8 protein expression in vitro in human induced pluripotent stem cell (hiPSC)-derived cell culture models. Methods A tetracysteine (TC) motif was introduced into the human MCT8 sequence at four different positions as binding sites for fluorescent biarsenical dyes. Human Embryonic Kidney 293 cells were transfected and stained with fluorescein-arsenical hairpin-binder (FlAsH). Counterstaining with specific MCT8 antibody was performed. Triiodothyronine (T3) uptake was indirectly measured with a T3 responsive luciferase-based reporter gene assay in Madin-Darby Canine Kidney 1 cells for functional characterization. Results FlAsH staining and antibody counterstaining of all four constructs showed cell membrane expression of all MCT8 constructs. The construct with the tag after the first start codon demonstrated comparable T3 uptake to the MCT8 wildtype. Conclusion Our data indicate that introduction of a TC-tag directly after the first start codon generates a MCT8 reporter with suitable characteristics for live-cell monitoring of MCT8 expression. One promising future application will be generation of stable hiPSC MCT8 reporter lines to characterize MCT8 expression patterns during in vitro neuronal development.

Live-Cell Probe for In Situ Single-Cell Monitoring of Mitochondrial DNA Methylation

ACS Sensors ◽  
2021 ◽  
Author(s):  
Han Zhao ◽  
Donghan Ma ◽  
Junkai Xie ◽  
Oscar Sanchez ◽  
Fang Huang ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Mitochondrial Dna ◽  
Single Cell ◽  
Live Cell ◽  
Cell Monitoring

scholarly journals LED-Based Tomographic Imaging for Live-Cell Monitoring of Pancreatic Islets in Microfluidic Channels

Proceedings ◽  
2017 ◽  
Vol 1 (4) ◽  
pp. 552 ◽  
Author(s):  
Gregor Scholz ◽  
Qifeng Xu ◽  
Torben Schulze ◽  
Heidi Boht ◽  
Kai Mattern ◽  
...  
Keyword(s):  
Pancreatic Islets ◽  
Tomographic Imaging ◽  
Live Cell ◽  
Microfluidic Channels ◽  
Cell Monitoring
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