Gene Delivery into Neuronal Cells by Calcium Phosphate-Mediated Transfection

Watson, Andrea; Latchman, David S.

doi:10.1006/meth.1996.0105

Cited by 18 publications

(9 citation statements)

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“…They were then cloned into the HpaI/ Xho I sites of a pSico vector (31), named pSico-miR-127. Viral particles were produced by calcium phosphate–mediated transfection into 293T cells as described (32). Lentiviral supernatants were collected 48 hours after transfection, passed through a 0.22-μm filter, and used directly to infect MH.S cells.…”

Section: Methodsmentioning

confidence: 99%

MicroRNA-127 Inhibits Lung Inflammation by Targeting IgG Fcγ Receptor I

Xie

Liang

Liu

et al. 2012

The Journal of Immunology

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The molecular mechanisms of acute lung injury are incompletely understood. MicroRNAs (miRNAs) are crucial biological regulators that act by suppressing their target genes and are involved in a variety of pathophysiologic processes. miR-127 appears to be down-regulated during lung injury. We set out to investigate the role of miR-127 in lung injury and inflammation. Expression of miR-127 significantly reduced cytokine release by macrophages. Looking into the mechanisms of regulation of inflammation by miR-127, we found that IgG Fcγ Receptor I (FcγRI/CD64) was a target of miR-127, as evidenced by reduced CD64 protein expression in macrophages over-expressing miR-127. Furthermore, miR-127 significantly reduced the luciferase activity with a reporter construct containing the native 3′-UTR of CD64. Importantly, we demonstrated that miR-127 attenuated lung inflammation in an IgG immune complex (IgG IC) model in vivo. Collectively, these data show that miR-127 targets macrophage CD64 expression and promotes the reduction of lung inflammation. Understanding how miRNAs regulate lung inflammation may represent an attractive way to control inflammation induced by infectious or non-infectious lung injury.

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Section: Methodsmentioning

confidence: 99%

MicroRNA-127 Inhibits Lung Inflammation by Targeting IgG Fcγ Receptor I

Xie

Liang

Liu

et al. 2012

The Journal of Immunology

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“…In that sense, nanosized CAP particles present one of the most viable options for gene delivery systems, and for that reason they have been extensively used as an in vitro gene delivery agent for over 30 years and are currently being investigated for in vivo purposes as well. 270,271 Structural and chemical features of CAP particles may provide all the essential requirements for their efficient usage as genetic transfection agents, including: the binding affinity of DNA molecules onto HAP crystals 272 ; the composite particle stability in extracellular space; an efficient cellular uptake and resorbability of the particles followed by a gradual release of the DNA and its escape from the endosomal network into the cytoplasmatic intracellular space, and eventual cytosolic transport and nuclear localization for transcription. 273 • Compared with other drug delivery carriers, CAPs possess the following advantages: • Favorable biodegradability and biocompatibility properties in general.…”

Section: Applicationmentioning

confidence: 99%

Nanosized hydroxyapatite and other calcium phosphates: Chemistry of formation and application as drug and gene delivery agents

2010

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The first part of this review looks at the fundamental properties of hydroxyapatite (HAP), the basic mineral constituent of mammalian hard tissues, including the physicochemical features that govern its formation by precipitation. A special emphasis is placed on the analysis of qualities of different methods of synthesis and of the phase transformations intrinsic to the formation of HAP following precipitation from aqueous solutions. This serves as an introduction to the second part and the main subject of this review, which relates to the discourse regarding the prospects of fabrication of ultrafine, nanosized particles based on calcium phosphate carriers with various therapeutic and/or diagnostic agents coated on and/or encapsulated within the particles. It is said that the particles could be either surface-functionalized with amphiphiles, peptides, proteins, or nucleic acids or injected with therapeutic agents, magnetic ions, or fluorescent molecules. Depending on the additive, they could be subsequently used for a variety of applications, including the controlled delivery and release of therapeutic agents (extracellularly or intracellularly), magnetic resonance imaging and hyperthermia therapy, cell separation, blood detoxification, peptide or oligonucleotide chromatography and ultrasensitive detection of biomolecules, and in vivo and in vitro gene transfection. Calcium phosphate nanoparticles as carriers of therapeutic agents that would enable a controlled drug release to treat a given bone infection and at the same be resorbed in the body so as to regenerate hard tissue lost to disease are emphasized hereby as one of the potentially attractive smart materials for the modern medicine.

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“…In vitro dual-luciferase reporter assay HEK293 cells were seeded in 24-well cell-culture plates at a density of 8 × 10 4 cells/well. After 18 h of growth, transfections were performed using a standard calcium-phosphate method according to established protocols (60)(61)(62). Each of the Firefly experimental luciferase vectors was co-transfected with a Renilla luciferase vector ( pRN-TK) (1 : 10 ratio of Renilla: experimental vector), which served as a transfection control.…”

Section: Cell Culturementioning

confidence: 99%

Genetic variants and cellular stressors associated with exfoliation syndrome modulate promoter activity of a lncRNA within theLOXL1locus

et al. 2015

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Exfoliation syndrome (XFS) is a common, age-related, systemic fibrillinopathy. It greatly increases risk of exfoliation glaucoma (XFG), a major worldwide cause of irreversible blindness. Coding variants in the lysyl oxidase-like 1 (LOXL1) gene are strongly associated with XFS in all studied populations, but a functional role for these variants has not been established. To identify additional candidate functional variants, we sequenced the entire LOXL1 genomic locus (∼40 kb) in 50 indigenous, black South African XFS cases and 50 matched controls. The variants with the strongest evidence of association were located in a welldefined 7-kb region bounded by the 3'-end of exon 1 and the adjacent region of intron 1 of LOXL1. We replicated this finding in US Caucasian (91 cases/1031 controls), German (771 cases/1365 controls) and Japanese (1484 cases/1188 controls) populations. The region of peak association lies upstream of LOXL1-AS1, a long non-coding RNA (lncRNA) encoded on the opposite strand of LOXL1. We show that this region contains a promoter and, importantly, that the strongly associated XFS risk alleles in the South African population are functional variants that significantly modulate the activity of this promoter. LOXL1-AS1 expression is also significantly altered in response to oxidative stress in human lens epithelial cells and in response to cyclic mechanical stress in human Schlemm's canal endothelial cells. Taken together, these findings support a functional role for the LOXL1-AS1 lncRNA in cellular stress response and suggest that dysregulation of its expression by genetic risk variants plays a key role in XFS pathogenesis.

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Gene Delivery into Neuronal Cells by Calcium Phosphate-Mediated Transfection

Cited by 18 publications

References 0 publications

MicroRNA-127 Inhibits Lung Inflammation by Targeting IgG Fcγ Receptor I

MicroRNA-127 Inhibits Lung Inflammation by Targeting IgG Fcγ Receptor I

Nanosized hydroxyapatite and other calcium phosphates: Chemistry of formation and application as drug and gene delivery agents

Genetic variants and cellular stressors associated with exfoliation syndrome modulate promoter activity of a lncRNA within theLOXL1locus

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