Visualizing the distribution and transport of mRNAs in living cells

Bratu, Diana P.; Cha, Byeong-Jik; Mhlanga, Musa M.; Kramer, Fred Russell; Tyagi, Sanjay

doi:10.1073/pnas.2233244100

Cited by 394 publications

(363 citation statements)

References 36 publications

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“…In addition, nanomaterials may protect DNA and facilitate DNA delivery into cells, [12][13][14] avoiding the use of transfection agents or microinjection. [15][16][17] Finally, several different probes can be attached to the same particle for multiplexed detection. 18,19 Recently, a number of fluorescent sensors have been prepared using carbon-based nanomaterials as quenchers (e.g.…”

Section: Introductionmentioning

confidence: 99%

Molecular Beacon Lighting up on Graphene Oxide

2012

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A molecular beacon (MB) is comprised of a fluorophore and a quencher linked by a DNA hairpin. MBs have been widely used for homogeneous DNA detection. In addition to molecular quenchers, many nanomaterials such as graphene oxide (GO) also possess excellent quenching efficiency. Most reported fluorescent sensors relied on DNA probes physisorbed by GO, which may suffer from non-specific probe displacement and false positive signal. In this work, we report the preparation and characterization of a MB using graphene oxide (GO) as quencher, where an amino and FAM (6carboxyfluorescein) dual labeled DNA was covalently attached to GO via an amide linkage. A major challenge was to remove noncovalently attached probes due to strong DNA adsorption by GO. While DNA desorption was favored at low salt, high pH, high temperature, and by using organic solvents, the complementary DNA was required to achieve complete desorption of non-covalently linked DNA probes. The DNA adsorption energy was measured using isothermal titration calorimetry, revealing the heterogeneous nature of GO. The covalent probe has a detection limit of 2.2 nM using a sample volume of 0.05 mL. With 2 mL sample, the detection limit can reach 150 pM. The covalent probe is highly resistant to non-specific probe displacement and will find applications in serum and cellular samples where high probe stability is demanded.

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Section: Introductionmentioning

confidence: 99%

Molecular Beacon Lighting up on Graphene Oxide

2012

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“…2; unpublished work) and has a profound effect on the distribution of germinal granules and the subsequent formation of germ cells in the embryos (Chan et al, , 2007. Further studies from our laboratory using molecular beacons technology (Bratu et al, 2003) showed that Xlsirts, VegT, and Fatvg RNA are integrated into the cytoskeleton in such a way that the individual RNAs have unique patterns of localization on cytokertain filaments ( Fig. 2; unpublished data).…”

Section: Structural Role Of Rna In the Cytoplasm Organization And Maimentioning

confidence: 91%

Teachings from the egg: New and unexpected functions of RNAs

Kloc

2009

Molecular Reproduction Devel

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SUMMARYLocalized mRNAs found in specific regions of somatic cells, germ cells, and embryos function through their protein translation products in cell polarization and development. Recent studies on Xenopus and Drosophila eggs and various somatic cells showed that some of the localized noncoding and coding RNAs play a structural (translation independent) role in maintaining the integrity of microtubule and microfilament cytoskeleton and/or may function in protein folding or as a scaffold for the assembly of cytoplasmic complexes essential for egg or embryo development. In addition, structural noncoding RNAs within the cell nucleus have been shown to be involved in the organization of chromatin, nuclear bodies, and DNA replication. The fact that some of the RNAs may have previously unforeseen structural functions, will change our view on traditional functions of RNAs and will open new frontiers in the field of RNA studies and therapeutic development.Mol. Reprod. Dev. 76: 922-932, 2009. ß

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“…mRNA localization in living cells is believed to be closely related to posttranscriptional regulation of gene expression but much remains to be seen if such localization indeed targets a protein to its site of function by producing the protein 'right on the spot'. Using a dual beacon approach, the transport and localization of Oskar mRNA in drosophila melanogaster oocytes has also been visualized [64]. In this work, molecular beacons with 2′-O-methyl backbone were delivered into cells using microinjection and the migration of Oskar mRNAs were tracked in real time, from the nurse cells where it is produced to the posterior cortex of the oocyte where it is localized.…”

Section: Molecular Beaconsmentioning

confidence: 99%

“…To label SPIOs with positron-emitting 64 Cu, the chelator 1,4,7,10-tetraazacyclo-dodecane-1,4,7,10-tetraacetic acid (DOTA) was conjugated to PEG termini. The 64 Cu-SPIO probes produced strong MR and PET signals, and were stable in mouse serum for 24 h at 37°C.…”

Section: Nanoparticle-based Mri and Mr/pet Imaging Contrast Agentsmentioning

confidence: 99%

“…The two imaging techniques are often combined with CT or MRI in order to locate the site of the signal. PET and SPECT utilize radioisotopes such as 64 Cu, 111 In and 99m Tc, the chelates of which can be conjugated to nanoparticles similar to that of Gd 3+ . For example, 64 Cu-DOTA was linked to siRNA, which is in turn conjugated to transferrin targeted polycation nanoparticles for combined cancer imaging and treatment [159].…”

Section: Nanoparticles For Combined Imaging and Therapymentioning

confidence: 99%

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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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Visualizing the distribution and transport of mRNAs in living cells

Cited by 394 publications

References 36 publications

Molecular Beacon Lighting up on Graphene Oxide

Molecular Beacon Lighting up on Graphene Oxide

Teachings from the egg: New and unexpected functions of RNAs

Engineering imaging probes and molecular machines for nanomedicine

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