pH-sensing nano-crystals of carbonate apatite: Effects on intracellular delivery and release of DNA for efficient expression into mammalian cells

<p>Gene therapy through intracellular delivery of a functional gene or a gene-silencing element is a promising approach to properly treat critical human diseases like cancer. The ability of synthetically designed small interfering RNA (siRNA) to effectively silence genes<em> </em>at post-transcriptional level has made them attractive options in targeted therapeutics. However, naked siRNA being unable to passively diffuse through cellular membranes, poses difficulty in fully exploiting the potential of the technology. pH-sensitive carbonate apatite has been developed as an efficient tool to deliver siRNA into the mammalian cells by virtue of its high affinity interaction with the siRNA and effective cellular endocytosis. Moreover, internalized siRNA has been found to escape from the endosomes in a time-dependent manner and effectively silenced reporter gene expression. Knockdown of cyclin B1 gene with only 10 nM of siRNA delivered by carbonate apatite has resulted in significant death of cervical cancer cells. Moreover, delivery of siRNA against cyclin B1 gene has led to the sensitization of the cancer cells to both cisplatin and doxorubicin at a particulat drug concentration. Thus, the new method of siRNA delivery is highly promising for pre-clinical and clinical cancer therapy using siRNA therapeutics.</p>

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Identification of types of membrane injuries and cell death using whole cell-based proton-sensitive field-effect transistor systems

The Analyst ◽

10.1039/c7an00502d ◽

2017 ◽

Vol 142 (18) ◽

pp. 3451-3458 ◽

Cited By ~ 5

Author(s):

Yuki Imaizumi ◽

Tatsuro Goda ◽

Akira Matsumoto ◽

Yuji Miyahara

Keyword(s):

Cell Death ◽

Mammalian Cells ◽

Field Effect ◽

Field Effect Transistor ◽

Ph Sensing ◽

Membrane Injury ◽

Whole Cell ◽

Sensing Systems ◽

Effect Transistor ◽

Chemical Stimuli

Membrane injury and apoptosis of mammalian cells by chemical stimuli were distinguished using ammonia-perfused continuous pH-sensing systems.

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Novel human-derived cell-penetrating peptides for specific subcellular delivery of therapeutic biomolecules

Biochemical Journal ◽

10.1042/bj20050401 ◽

2005 ◽

Vol 390 (2) ◽

pp. 407-418 ◽

Cited By ~ 83

Author(s):

Catherine de Coupade ◽

Antonio Fittipaldi ◽

Vanessa Chagnas ◽

Matthieu Michel ◽

Sophie Carlier ◽

...

Keyword(s):

Cell Membrane ◽

Mammalian Cells ◽

Heparan Sulphate ◽

Intracellular Delivery ◽

Membrane Lipid ◽

Cell Penetrating Peptides ◽

Heparin Binding ◽

Independent Manner ◽

Cell Penetrating

Short peptide sequences that are able to transport molecules across the cell membrane have been developed as tools for intracellular delivery of therapeutic molecules. This work describes a novel family of cell-penetrating peptides named Vectocell® peptides [also termed DPVs (Diatos peptide vectors)]. These peptides, originating from human heparin binding proteins and/or anti-DNA antibodies, once conjugated to a therapeutic molecule, can deliver the molecule to either the cytoplasm or the nucleus of mammalian cells. Vectocell® peptides can drive intracellular delivery of molecules of varying molecular mass, including full-length active immunoglobulins, with efficiency often greater than that of the well-characterized cell-penetrating peptide Tat. The internalization of Vectocell® peptides has been demonstrated to occur in both adherent and suspension cell lines as well as in primary cells through an energy-dependent endocytosis process, involving cell-membrane lipid rafts. This endocytosis occurs after binding of the cell-penetrating peptides to extracellular heparan sulphate proteoglycans, except for one particular peptide (DPV1047) that partially originates from an anti-DNA antibody and is internalized in a caveolar independent manner. These new therapeutic tools are currently being developed for intracellular delivery of a number of active molecules and their potentiality for in vivo transduction investigated.

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pH-Sensitive Nanocrystals of Carbonate Apatite- a Powerful and Versatile Tool for Efficient Delivery of Genetic Materials to Mammalian Cells

Advances in Biomaterials Science and Biomedical Applications ◽

10.5772/53107 ◽

2013 ◽

Author(s):

Ezharul Hoque

Keyword(s):

Mammalian Cells ◽

Ph Sensitive ◽

Carbonate Apatite ◽

Efficient Delivery ◽

Versatile Tool

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Functional genetic encoding of sulfotyrosine in mammalian cells

Nature Communications ◽

10.1038/s41467-020-18629-9 ◽

2020 ◽

Vol 11 (1) ◽

Author(s):

Xinyuan He ◽

Yan Chen ◽

Daisy Guiza Beltran ◽

Maia Kelly ◽

Bin Ma ◽

...

Keyword(s):

Mammalian Cells ◽

Biochemical Characterization ◽

Trna Synthetase ◽

Functional Verification ◽

Cellular Processes ◽

Genetic Encoding ◽

Efficient Expression

Abstract Protein tyrosine O-sulfation (PTS) plays a crucial role in extracellular biomolecular interactions that dictate various cellular processes. It also involves in the development of many human diseases. Regardless of recent progress, our current understanding of PTS is still in its infancy. To promote and facilitate relevant studies, a generally applicable method is needed to enable efficient expression of sulfoproteins with defined sulfation sites in live mammalian cells. Here we report the engineering, in vitro biochemical characterization, structural study, and in vivo functional verification of a tyrosyl-tRNA synthetase mutant for the genetic encoding of sulfotyrosine in mammalian cells. We further apply this chemical biology tool to cell-based studies on the role of a sulfation site in the activation of chemokine receptor CXCR4 by its ligand. Our work will not only facilitate cellular studies of PTS, but also paves the way for economical production of sulfated proteins as therapeutic agents in mammalian systems.

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Intracellular delivery of NF-κB small interfering RNA for modulating therapeutic activities of classical anti-cancer drugs in human cervical cancer cells

Drugs and Therapy Studies ◽

10.4081/dts.2013.e7 ◽

2013 ◽

Vol 3 (1) ◽

pp. 7 ◽

Cited By ~ 1

Author(s):

Anthony Stanislaus ◽

Anil Philip Kunnath ◽

Snigdha Tiash ◽

Tahereh Fatemian ◽

Nur Izyani Kamaruzman ◽

...

Keyword(s):

Cervical Cancer ◽

Cancer Cells ◽

Intracellular Delivery ◽

Cyclin B1 ◽

Carbonate Apatite ◽

Cancer Drugs ◽

Small Interfering Rnas ◽

Strong Electrostatic Interaction ◽

Anti Cancer ◽

Gene Transcripts

Cervical cancer is the second most common cancer and fourth leading cause of cancer-related deaths among women. Advanced stage of the disease is treated with radiation therapy and chemotherapy with poor therapeutic outcome and adverse side effects. NFκB, a well-known transcription factor in the control of immunity and inflammation, has recently emerged as a key regulator of cell survival through induction of antiapoptotic genes. Many human cancers, including cervical carcinoma, constitutively express NF-κB and a blockade in expression of its subunit proteins through targeted knockdown of the gene transcripts with small interfering RNAs (siRNA) could be an attractive approach in order to sensitize the cancer cells towards the widely used anti-cancer drugs. However, the inefficiency of the naked siRNA to cross the plasma membrane and its sensitiveness to nuclease-mediated degradation are the major challenges limiting the siRNA technology in therapeutic intervention. pH-sensitive carbonate apatite has been established as an efficient nano-carrier for intracellular delivery of siRNA, due to its strong electrostatic interaction with the siRNA, the desirable size distribution of the resulting siRNA complex for effective endocytosis and the ability of the endocytosed siRNA to be released from the degradable particles and escape the endosomes, thus leading to the effective knockdown of the target gene of cyclin B1 or ABCB1. Here, we report that carbonate apatite-facilitated delivery of the siRNA targeting NF-κB1 and NF-κB2 gene transcripts in HeLa, a human cervical adenocar- cinoma cell line expressing NF-κB, led to a synergistic effect in enhancement of chemosensitivity to doxorubicin, but apparently not to cisplatin or paclitaxel.

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Synthesis, Cellular Uptake, and Cytotoxicity of a Thermally Responsive Nanocapsule

ASME 2009 Summer Bioengineering Conference, Parts A and B ◽

10.1115/sbc2009-206872 ◽

2009 ◽

Author(s):

Wujie Zhang ◽

Jianhua Rong ◽

Qian Wang ◽

Xiaoming He

Keyword(s):

Cellular Uptake ◽

Blood Cells ◽

Mammalian Cells ◽

Polymeric Nanoparticles ◽

Intracellular Delivery ◽

Sustained Delivery ◽

Hydrogel Nanoparticles ◽

Ambient Temperatures ◽

Thermally Responsive ◽

Therapeutic Drugs

Recently, polymeric nanoparticles have attracted tremendous interests as a useful tool to encapsulate therapeutic drugs, genes, and proteins for their controlled and sustained delivery. Among them, polymeric hydrogel nanoparticles with thermal and/or pH responsiveness have attracted particular attention [1]. Trehalose, a non-reducing disaccharide of glucose, has been demonstrated to be a potent, nontoxic bioprotectant for stabilizing lipids, proteins, viruses, and blood cells at cryogenic and particularly, ambient temperatures (i.e., cryo and lyopreservation) [2]. However, intracellular delivery of trehalose into small eukaryotic mammalian cells in a large quantity for biostabilization purpose has not been very successful so far [2]. In this study, a thermally responsive polymeric nanocapsule was synthesized and characterized with the aim to encapsulate trehalose for its intracellular delivery.

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