High-Throughput Tracking of Pluripotent Human Embryonic Stem Cells with Dual Fluorescence Resonance Energy Transfer Molecular Beacons

King, Frank W.; Liszewski, Walter; Ritner, Carissa; Bernstein, Harold S.

doi:10.1089/scd.2010.0219

Cited by 39 publications

(37 citation statements)

References 23 publications

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“…The reagent (4-10 mg) was incubated with 120 mM CaCl 2 and HEPES buffer containing NaCl, NaHPO 4 , and HEPES, pH 7.1, for 10 minutes on ice. The mixture was immediately added to the surface of the cultured cells, swirled gently, and returned to the incubator at 37°C with 5% CO 2 until needed.…”

Section: Calcium Phosphate Precipitationmentioning

confidence: 99%

“…Although RNA beacons can be developed to detect genes associated with pluripotency (e.g., Oct4, nanog, Sox2) [1,4], many stem cell applications create induced pluripotent stem cells and induce them to differentiate toward specific lineages [5] by introducing specific genes (often transcription factors). Current selection methods require a distinctive surface marker that can detect target cells using antibodies or require introduction of a reporter gene such as green fluorescent protein (GFP).…”

Section: Introductionmentioning

confidence: 99%

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Development of a Universal RNA Beacon for Exogenous Gene Detection

Guo

Cohen

et al. 2015

Stem Cells Translational Medicine

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Stem cell therapy requires a nontoxic and high-throughput method to achieve a pure cell population to prevent teratomas that can occur if even one cell in the implant has not been transformed. A promising method to detect and separate cells expressing a particular gene is RNA beacon technology. However, developing a successful, specific beacon to a particular transfected gene can take months to develop and in some cases is impossible. Here, we report on an off-the-shelf universal beacon that decreases the time and cost of applying beacon technology to select any living cell population transfected with an exogenous gene. STEM CELLS TRANSLATIONAL MEDICINE 2015;4:476-482 SIGNIFICANCEStem cell therapy requires pure cell populations. RNA beacons have a high potential to select and separate differentiated cells from undifferentiated cells. However, development of a beacon for a specific gene can be difficult and time-consuming. An RNA tag was developed that can be attached to any gene without affecting the protein product. The tagged gene is detected by an off-the-shelf RNA beacon that can be used for cell selection.

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Section: Calcium Phosphate Precipitationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Development of a Universal RNA Beacon for Exogenous Gene Detection

Guo

Cohen

et al. 2015

Stem Cells Translational Medicine

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“…adapted dual-fluorescence resonance energy transfer "molecular beacon" technology for transient, real-time detection of gene expression during hESC differentiation (62). Molecular beacons are single-stranded oligonucleotide probes that have been employed to assay gene expression in vitro, as in realtime PCR, and in vivo using microscopy.…”

Section: Reviewmentioning

confidence: 99%

“…Molecular beacons are single-stranded oligonucleotide probes that have been employed to assay gene expression in vitro, as in realtime PCR, and in vivo using microscopy. These investigators showed that appropriately designed, dual-fluorescence resonance energy transfer molecular beacon pairs can identify the expression of specific mRNAs and facilitate the collection of specific hESC populations by fluorescence-activated cell sorting, while leaving the hESC genome intact (62).…”

Section: Reviewmentioning

confidence: 99%

Stem cell therapy for cardiac disease

2012

Self Cite

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“…Dual FRET molecular beacons have been used to detect K-Ras, surviving and oct4 mRNAs in HDF, Miapaca-2, and H1 cells, respectively [66,72]. Interestingly, k-Ras mRNAs seemed to localize with a filamentous pattern in HDF cell whereas survivin mRNAs seemed to localize in a non-symmetrical pattern within Miapaca-2 cells, often to one side of the nucleus of the cell.…”

Section: Molecular Beaconsmentioning

confidence: 99%

Engineering imaging probes and molecular machines for nanomedicine

Tong

Cradick

et al. 2012

Sci. China Life Sci.

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Nanomedicine is an emerging field that integrates nanotechnology, biomolecular engineering, life sciences and medicine; it is expected to produce major breakthroughs in medical diagnostics and therapeutics. Due to the size-compatibility of nano-scale structures and devices with proteins and nucleic acids, the design, synthesis and application of nanoprobes, nanocarriers and nanomachines provide unprecedented opportunities for achieving a better control of biological processes, and drastic improvements in disease detection, therapy, and prevention. Recent advances in nanomedicine include the development of functional nanoparticle based molecular imaging probes, nano-structured materials as drug/gene carriers for in vivo delivery, and engineered molecular machines for treating single-gene disorders. This review focuses on the development of molecular imaging probes and engineered nucleases for nanomedicine, including quantum dot bioconjugates, quantum dot-fluorescent protein FRET probes, molecular beacons, magnetic and gold nanoparticle based imaging contrast agents, and the design and validation of zinc finger nucleases (ZFNs) and TAL effector nucleases (TALENs) for gene targeting. The challenges in translating nanomedicine approaches to clinical applications are discussed. Nanomedicine is an emerging field that integrates nanotechnology, biomolecular engineering, biology, and medicine [1]. It focuses on the development of engineered nano-scale (1100 nm) materials, structures and devices for better diagnostics and highly specific medical intervention in curing disease or repairing damaged tissues. As a basis for nanomedicine, nanotechnology is the science, engineering, and technology related to the understanding and control of matter at the nano-scale, and the development of materials, devices, and systems that have novel properties and functions due to their nano-scale dimensions or components. Nanotechnology also provides new abilities to measure, control and manipulate matter (including soft matter) at the nano-scale what was unthinkable with conventional tools. Owing to the size-compatibility of nano-scale structures with proteins and nucleic acids in living cells, nanomedicine approaches have the potential to provide unprecedented opportunities for achieving a better control of biological processes, and drastic improvements in disease detection, therapy, and prevention, thus revolutionizing medicine. Over the last ten years or so, significant efforts have been made in the US, China, Europe and elsewhere to develop nanomedicine. For example, the US National Institutes of Health has developed a nanomedicine centers network, and invested a significant amount of research funding to nanomedicine development. Just in FY 2009 (Oct. 1, 2008Sept. 30, 2009, the total NIH funding in nanotechnology/ nanoscience projects was more than 410 million US dollars. SPECIAL TOPIC

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High-Throughput Tracking of Pluripotent Human Embryonic Stem Cells with Dual Fluorescence Resonance Energy Transfer Molecular Beacons

Cited by 39 publications

References 23 publications

Development of a Universal RNA Beacon for Exogenous Gene Detection

Development of a Universal RNA Beacon for Exogenous Gene Detection

Stem cell therapy for cardiac disease

Engineering imaging probes and molecular machines for nanomedicine

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