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RNA mimics of green fluorescent protein.
Science. 2011 Jul 29; 333(6042):642-6
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01 Aug 2011
New Finding, Technical Advance
DOI: 10.3410/f.12351957.13537055
Green fluorescent protein (GFP) and related constructs have become widely used for subcellular localisation of tagged proteins in cells. This article describes a new technique that may prove to be of equal value to many researchers -- a way to fluorescently tag specific RNAs for localisation in vivo.
Not...Continue reading
Not...Continue reading
Green fluorescent protein (GFP) and related constructs have become widely used for subcellular localisation of tagged proteins in cells. This article describes a new technique that may prove to be of equal value to many researchers -- a way to fluorescently tag specific RNAs for localisation in vivo.
Not only do the authors prove the principle of the approach, they demonstrate its application to a specific RNA in mammalian cells and develop a range of tag-fluorophore combinations to allow fluorescence at varying wavelengths. To date, subcellular localisation of RNAs has proven difficult -- this new approach promises rapid advances in our understanding of the dynamics of RNA localisation in the cell.
Not only do the authors prove the principle of the approach, they demonstrate its application to a specific RNA in mammalian cells and develop a range of tag-fluorophore combinations to allow fluorescence at varying wavelengths. To date, subcellular localisation of RNAs has proven difficult -- this new approach promises rapid advances in our understanding of the dynamics of RNA localisation in the cell.
Disclosures
None declared
Add Comment
Clark C: F1000Prime Recommendation of [Paige JS et al., Science 2011, 333(6042):642-6]. In F1000Prime, 01 Aug 2011; DOI: 10.3410/f.12351957.13537055. F1000Prime.com/12351957#eval13537055
08 Aug 2011
New Finding
DOI: 10.3410/f.12351957.13747058
This work is important because Paige and co-workers have provided a simple method to fluorescently tag an RNA molecule within cells. The ability to confer green fluorescent protein (GFP)-like functionality to RNA molecules will allow studies of the synthesis and fate of tagged RNAs in ways that will most...Continue reading
This work is important because Paige and co-workers have provided a simple method to fluorescently tag an RNA molecule within cells. The ability to confer green fluorescent protein (GFP)-like functionality to RNA molecules will allow studies of the synthesis and fate of tagged RNAs in ways that will most likely redefine our understanding of many aspects of RNA biology, particularly processes involving RNA dynamics, which, in the past, had been limited to studies employing fixed cells or cell extracts.
GFP has become an excellent tool to study many different aspects of biology, including processes such as protein folding, protein transport, and RNA-protein dynamics. The ability of GFP to form an intracellular chromophore that exhibits fluorescence in the presence of molecular oxygen has transformed the way we visualize molecules -- especially proteins -- in living cells. We can now track when and where an experimentally designed GFP-fusion protein is made and how it moves inside cells. However, the same was not as easily accomplished for RNA.
Now, Paige and co-workers have simplified the imaging of RNAs in living cells. To do so, they used artificial evolution (SELEX) to select for RNA sequences (RNA aptamers) that manifest GFP-like properties when bound to small molecules (fluorophores) that resemble the fluorophore in GFP. Since the exhibition of fluorescence requires an RNA aptamer, cellular RNAs do not interfere with the analyses.
Amazingly, further rounds of selection identified additional RNA aptamer-fluorophore complexes with distinct spectra in the cyan, yellow, greenish-yellow or orange-red range. One RNA aptamer was dubbed 'Spinach' because of its enhanced GFP-like properties. The Spinach-fluorophore complex exhibited strong fluorescence when expressed in living bacterial cells. That the selected RNA aptamers could also be useful tools for tagging RNAs in living mammalian cells was demonstrated by fusing Spinach to the 3' end of the small non-coding 5S ribosomal (r)RNA. Spinach-5S rRNA fluorescence could be detected throughout the cell and manifested distribution similar to that of endogenous 5S rRNA. Furthermore, the dynamics of Spinach-5S rRNA fluorescence after the induction of stress granule formation by sucrose treatment or after inhibiting RNA transport using leptomycin-B recapitulated the known behavior of endogenous 5SrRNA.
GFP has become an excellent tool to study many different aspects of biology, including processes such as protein folding, protein transport, and RNA-protein dynamics. The ability of GFP to form an intracellular chromophore that exhibits fluorescence in the presence of molecular oxygen has transformed the way we visualize molecules -- especially proteins -- in living cells. We can now track when and where an experimentally designed GFP-fusion protein is made and how it moves inside cells. However, the same was not as easily accomplished for RNA.
Now, Paige and co-workers have simplified the imaging of RNAs in living cells. To do so, they used artificial evolution (SELEX) to select for RNA sequences (RNA aptamers) that manifest GFP-like properties when bound to small molecules (fluorophores) that resemble the fluorophore in GFP. Since the exhibition of fluorescence requires an RNA aptamer, cellular RNAs do not interfere with the analyses.
Amazingly, further rounds of selection identified additional RNA aptamer-fluorophore complexes with distinct spectra in the cyan, yellow, greenish-yellow or orange-red range. One RNA aptamer was dubbed 'Spinach' because of its enhanced GFP-like properties. The Spinach-fluorophore complex exhibited strong fluorescence when expressed in living bacterial cells. That the selected RNA aptamers could also be useful tools for tagging RNAs in living mammalian cells was demonstrated by fusing Spinach to the 3' end of the small non-coding 5S ribosomal (r)RNA. Spinach-5S rRNA fluorescence could be detected throughout the cell and manifested distribution similar to that of endogenous 5S rRNA. Furthermore, the dynamics of Spinach-5S rRNA fluorescence after the induction of stress granule formation by sucrose treatment or after inhibiting RNA transport using leptomycin-B recapitulated the known behavior of endogenous 5SrRNA.
Disclosures
None declared
Maquat L and Isken O: F1000Prime Recommendation of [Paige JS et al., Science 2011, 333(6042):642-6]. In F1000Prime, 08 Aug 2011; DOI: 10.3410/f.12351957.13747058. F1000Prime.com/12351957#eval13747058
10 Aug 2011
New Finding
DOI: 10.3410/f.12351957.13814054
In this work, the authors report a new fluorescent RNA aptamer with important imaging applications to RNA processes.
Fluorescent biomolecules have long been used as important tools for bio-imaging to visualize various cellular processes. The green fluorescent protein (GFP), first isolated from jell has...Continue reading
Fluorescent biomolecules have long been used as important tools for bio-imaging to visualize various cellular processes. The green fluorescent protein (GFP), first isolated from jell has...Continue reading
In this work, the authors report a new fluorescent RNA aptamer with important imaging applications to RNA processes.
Fluorescent biomolecules have long been used as important tools for bio-imaging to visualize various cellular processes. The green fluorescent protein (GFP), first isolated from jellyfish, has been used for numerous applications. The fluorescence mechanism of GFP is well understood in terms of specific contacts formed with fluorophores upon GFP folding; related fluorescent proteins have been developed. Similar bio-imaging techniques for RNA processes have also been sought, resulting, for example, in an RNA aptamer termed MGA that binds to the small industrial dye malachite green (MG). However, such fluorescent RNAs generally lack specificity in their activation mechanism and/or exhibit cytotoxicity. To remedy this, Paige et al. report the successful design of an RNA-fluorophore complex termed Spinach, which is both specific and robust, displaying a wide range of colors with functionally relevant brightness, and extensibility with other RNAs. This 24-residue RNA aptamer was designed based on GFP using the SELEX (systematic evolution of ligands by exponential enrichment) technology. Spinach has the potential for live-cell imaging of varied RNA processes.
Fluorescent biomolecules have long been used as important tools for bio-imaging to visualize various cellular processes. The green fluorescent protein (GFP), first isolated from jellyfish, has been used for numerous applications. The fluorescence mechanism of GFP is well understood in terms of specific contacts formed with fluorophores upon GFP folding; related fluorescent proteins have been developed. Similar bio-imaging techniques for RNA processes have also been sought, resulting, for example, in an RNA aptamer termed MGA that binds to the small industrial dye malachite green (MG). However, such fluorescent RNAs generally lack specificity in their activation mechanism and/or exhibit cytotoxicity. To remedy this, Paige et al. report the successful design of an RNA-fluorophore complex termed Spinach, which is both specific and robust, displaying a wide range of colors with functionally relevant brightness, and extensibility with other RNAs. This 24-residue RNA aptamer was designed based on GFP using the SELEX (systematic evolution of ligands by exponential enrichment) technology. Spinach has the potential for live-cell imaging of varied RNA processes.
Disclosures
None declared
Schlick T: F1000Prime Recommendation of [Paige JS et al., Science 2011, 333(6042):642-6]. In F1000Prime, 10 Aug 2011; DOI: 10.3410/f.12351957.13814054. F1000Prime.com/12351957#eval13814054
11 Aug 2011
New Finding, Technical Advance
DOI: 10.3410/f.12351957.13837064
We think this article is exciting and important because the authors discovered RNA aptamers that cause green fluorescent protein (GFP)-like fluorescence when they bind to small fluorophores and demonstrated their use for live-cell imaging of RNA. There is enormous potential for using this technology to...Continue reading
We think this article is exciting and important because the authors discovered RNA aptamers that cause green fluorescent protein (GFP)-like fluorescence when they bind to small fluorophores and demonstrated their use for live-cell imaging of RNA. There is enormous potential for using this technology to study RNA biology.
Different aptamers and different fluorophores in combination generate fluorescence emission peaks in different regions of the visible spectrum. By fusing RNA aptamers to 5S ribosomal RNA (rRNA) sequences, and expressing the fusion genes in cells, the application of these RNA-fluorophore complexes for live-cell imaging of RNA was demonstrated.
Different aptamers and different fluorophores in combination generate fluorescence emission peaks in different regions of the visible spectrum. By fusing RNA aptamers to 5S ribosomal RNA (rRNA) sequences, and expressing the fusion genes in cells, the application of these RNA-fluorophore complexes for live-cell imaging of RNA was demonstrated.
Disclosures
None declared
Sachs M and Lin X: F1000Prime Recommendation of [Paige JS et al., Science 2011, 333(6042):642-6]. In F1000Prime, 11 Aug 2011; DOI: 10.3410/f.12351957.13837064. F1000Prime.com/12351957#eval13837064
11 Aug 2011
Technical Advance
DOI: 10.3410/f.12351957.13860055
This paper describes what will undoubtedly prove to be a useful tool for studying RNA biology. From combinatorial libraries, the authors selected RNAs that bind to organic compounds that closely resemble the chromophore in green fluorescent protein (GFP). The molecule is not fluorescent in its free...Continue reading
This paper describes what will undoubtedly prove to be a useful tool for studying RNA biology. From combinatorial libraries, the authors selected RNAs that bind to organic compounds that closely resemble the chromophore in green fluorescent protein (GFP). The molecule is not fluorescent in its free form, but becomes fluorescent when bound by the RNAs. Different RNAs produce different colored complexes with excellent brightness, comparable to that of GFP.
Moreover, cellular RNAs could be tagged with these sequences and, upon addition of the pro-fluorescent small molecule, the resultant complex could be used to image the localization of the expressed RNA. A nice feature of these RNA-small molecule complexes is that they appear to be quite resistant to photobleaching.
This technology will facilitate cell biological studies of RNA localization, lifetime and transport.
Moreover, cellular RNAs could be tagged with these sequences and, upon addition of the pro-fluorescent small molecule, the resultant complex could be used to image the localization of the expressed RNA. A nice feature of these RNA-small molecule complexes is that they appear to be quite resistant to photobleaching.
This technology will facilitate cell biological studies of RNA localization, lifetime and transport.
Disclosures
None declared
Kodadek T: F1000Prime Recommendation of [Paige JS et al., Science 2011, 333(6042):642-6]. In F1000Prime, 11 Aug 2011; DOI: 10.3410/f.12351957.13860055. F1000Prime.com/12351957#eval13860055
22 Aug 2011
New Finding, Technical Advance
DOI: 10.3410/f.12351957.14059054
A new molecular tool illuminates the RNA world.
In this study, the authors have used the SELEX (systematic evolution of ligands by exponential enrichment) technology to fluorescently tag RNA molecules. The RNA-fluorophore complex, named Spinach, developed by this method emits green fluorescence that...Continue reading
In this study, the authors have used the SELEX (systematic evolution of ligands by exponential enrichment) technology to fluorescently tag RNA molecules. The RNA-fluorophore complex, named Spinach, developed by this method emits green fluorescence that...Continue reading
A new molecular tool illuminates the RNA world.
In this study, the authors have used the SELEX (systematic evolution of ligands by exponential enrichment) technology to fluorescently tag RNA molecules. The RNA-fluorophore complex, named Spinach, developed by this method emits green fluorescence that is comparable to GFPs (green florescent proteins) and is markedly resistant to photobleaching. These properties make Spinach a highly useful molecular tool to study RNA processing and trafficking in live cells.
In this study, the authors have used the SELEX (systematic evolution of ligands by exponential enrichment) technology to fluorescently tag RNA molecules. The RNA-fluorophore complex, named Spinach, developed by this method emits green fluorescence that is comparable to GFPs (green florescent proteins) and is markedly resistant to photobleaching. These properties make Spinach a highly useful molecular tool to study RNA processing and trafficking in live cells.
Disclosures
None declared
Dutta A and Karnani N: F1000Prime Recommendation of [Paige JS et al., Science 2011, 333(6042):642-6]. In F1000Prime, 22 Aug 2011; DOI: 10.3410/f.12351957.14059054. F1000Prime.com/12351957#eval14059054
24 Aug 2011
New Finding, Technical Advance
DOI: 10.3410/f.12351957.14094054
This paper describes a powerful new method for fluorescent tagging of specific RNAs. The authors have developed RNA aptamers that bind to fluorophores with similar structures to the fluorophore in green fluorescent protein (GFP). Importantly, these fluorophores have no emission in the absence of aptamer...Continue reading
This paper describes a powerful new method for fluorescent tagging of specific RNAs. The authors have developed RNA aptamers that bind to fluorophores with similar structures to the fluorophore in green fluorescent protein (GFP). Importantly, these fluorophores have no emission in the absence of aptamer binding. This exciting technology will enable a range of applications, including in vivo tracking of RNA localization, RNA-RNA binding and RNA-protein binding.
Several methods have been developed for detecting specific RNA localization in cells, but all involve the co-expression of a target RNA plus a fluorescent protein that binds to specific sites in the RNA. This can be problematic because the unbound protein is also fluorescent and can obscure the signal. In addition, both components have to be expressed at low levels, which often cannot be easily controlled. This new technology requires only the expression of an RNA with the added aptamer sequence. The fluorophores are cell permeant, and are non-toxic and non-fluorescent in the absence of aptamer. Importantly, the authors have developed a range of fluorophores that cover the visible spectrum, which should enable the simultaneous tracking of multiple RNAs.
Several methods have been developed for detecting specific RNA localization in cells, but all involve the co-expression of a target RNA plus a fluorescent protein that binds to specific sites in the RNA. This can be problematic because the unbound protein is also fluorescent and can obscure the signal. In addition, both components have to be expressed at low levels, which often cannot be easily controlled. This new technology requires only the expression of an RNA with the added aptamer sequence. The fluorophores are cell permeant, and are non-toxic and non-fluorescent in the absence of aptamer. Importantly, the authors have developed a range of fluorophores that cover the visible spectrum, which should enable the simultaneous tracking of multiple RNAs.
Disclosures
None declared
Macara I: F1000Prime Recommendation of [Paige JS et al., Science 2011, 333(6042):642-6]. In F1000Prime, 24 Aug 2011; DOI: 10.3410/f.12351957.14094054. F1000Prime.com/12351957#eval14094054
01 Sep 2011
New Finding, Technical Advance
DOI: 10.3410/f.12351957.14379054
This paper describes an impressive technical advance that is likely to be a significant new tool in our ability to image RNAs in the cell.
Disclosures
None declared
Batey R: F1000Prime Recommendation of [Paige JS et al., Science 2011, 333(6042):642-6]. In F1000Prime, 01 Sep 2011; DOI: 10.3410/f.12351957.14379054. F1000Prime.com/12351957#eval14379054
08 Sep 2011
New Finding, Technical Advance
DOI: 10.3410/f.12351957.14452062
The study of protein trafficking was exponentially advanced by the development of green fluorescent protein (GFP) fusion protein technology. This exciting work by Paige et al. suggests that a similar advance in the study of cellular RNAs may now be possible. The work may also have significant implications...Continue reading
The study of protein trafficking was exponentially advanced by the development of green fluorescent protein (GFP) fusion protein technology. This exciting work by Paige et al. suggests that a similar advance in the study of cellular RNAs may now be possible. The work may also have significant implications for the study of RNA virus biology including virus entry, viral RNA replication, RNA trafficking and packaging, and innate systems of anti-viral immunity.
This paper describes a novel tool for visualization of RNAs in live cells. Using the systematic evolution of ligands by exponential enrichment (SELEX) methodology, RNA aptamers were selected that induce a robust fluorescent signal when bound to a GFP-like fluorophore. To demonstrate the utility of the system, one aptamer called 'Spinach' was fused to the 3' end of 5S, a small non-coding ribosomal RNA that normally associates with the large ribosomal subunit. The Spinach-5S complex was shown to localize within HEK 293T cells in a distribution similar to that of untagged 5S rRNA.
This paper describes a novel tool for visualization of RNAs in live cells. Using the systematic evolution of ligands by exponential enrichment (SELEX) methodology, RNA aptamers were selected that induce a robust fluorescent signal when bound to a GFP-like fluorophore. To demonstrate the utility of the system, one aptamer called 'Spinach' was fused to the 3' end of 5S, a small non-coding ribosomal RNA that normally associates with the large ribosomal subunit. The Spinach-5S complex was shown to localize within HEK 293T cells in a distribution similar to that of untagged 5S rRNA.
Disclosures
None declared
Garcia-Sastre A and Wang T: F1000Prime Recommendation of [Paige JS et al., Science 2011, 333(6042):642-6]. In F1000Prime, 08 Sep 2011; DOI: 10.3410/f.12351957.14452062. F1000Prime.com/12351957#eval14452062
15 Sep 2011
Zhihua Li
F1000 Genomics & Genetics
University of Pittsburgh School of Medicine,
Pittsburgh, PA, USA.
F1000 Genomics & Genetics
University of Pittsburgh School of Medicine,
Pittsburgh, PA, USA.
New Finding, Technical Advance
DOI: 10.3410/f.12351957.14567055
This fantastic work provides a new tool for imaging RNA molecules in living cells.
Fluorescent proteins are widely used to image proteins inside a living cell. However, imaging RNA molecules in a living cell is not that simple. One common technique involves tagging RNA molecules with stem-loop elements...Continue reading
Fluorescent proteins are widely used to image proteins inside a living cell. However, imaging RNA molecules in a living cell is not that simple. One common technique involves tagging RNA molecules with stem-loop elements...Continue reading
This fantastic work provides a new tool for imaging RNA molecules in living cells.
Fluorescent proteins are widely used to image proteins inside a living cell. However, imaging RNA molecules in a living cell is not that simple. One common technique involves tagging RNA molecules with stem-loop elements that specifically bind to fusion protein MS2-green fluorescent protein (GFP) (MS2 is a phage capsid protein), such that the RNA molecules are indirectly visualized by GFP. In this paper, Jeremy et al. took an alternative approach to lighting up RNA molecules inside the cell. They took advantage of the nature-designed fluorophore for GFP and selected several aptamers that can bind to GFP-like fluorophores to emit fluorescence. A wide range of spectra were generated by combining different aptamers with GFP-like fluorophores. In particular, after binding to an enhanced GFP (EGFP)-like fluorophore, one aptamer termed 'Spinach' can emit bright green fluorescence comparable to GFP. A great advantage by using GFP-like fluorophores is that they exhibit much less cytotoxicity compared to a previously used fluorophore, malachite green. This new tool will be extremely useful for studying RNA molecules using imaging technology. In the future, the aptamers for other spectra, such as blue and red, might be improved to increase their brightness for multiplex imaging.
Fluorescent proteins are widely used to image proteins inside a living cell. However, imaging RNA molecules in a living cell is not that simple. One common technique involves tagging RNA molecules with stem-loop elements that specifically bind to fusion protein MS2-green fluorescent protein (GFP) (MS2 is a phage capsid protein), such that the RNA molecules are indirectly visualized by GFP. In this paper, Jeremy et al. took an alternative approach to lighting up RNA molecules inside the cell. They took advantage of the nature-designed fluorophore for GFP and selected several aptamers that can bind to GFP-like fluorophores to emit fluorescence. A wide range of spectra were generated by combining different aptamers with GFP-like fluorophores. In particular, after binding to an enhanced GFP (EGFP)-like fluorophore, one aptamer termed 'Spinach' can emit bright green fluorescence comparable to GFP. A great advantage by using GFP-like fluorophores is that they exhibit much less cytotoxicity compared to a previously used fluorophore, malachite green. This new tool will be extremely useful for studying RNA molecules using imaging technology. In the future, the aptamers for other spectra, such as blue and red, might be improved to increase their brightness for multiplex imaging.
Disclosures
None declared
Marcotte E and Li Z: F1000Prime Recommendation of [Paige JS et al., Science 2011, 333(6042):642-6]. In F1000Prime, 15 Sep 2011; DOI: 10.3410/f.12351957.14567055. F1000Prime.com/12351957#eval14567055
04 Oct 2011
New Finding
DOI: 10.3410/f.12351957.14692101
We fully endorse the current evaluations of this paper. The enhanced green fluorescent protein (eGFP)-like RNA aptamer presented by the authors promises to open new avenues in the field of RNA live imaging. We are now excited to see the first applications of this new technique....Continue reading
We fully endorse the current evaluations of this paper. The enhanced green fluorescent protein (eGFP)-like RNA aptamer presented by the authors promises to open new avenues in the field of RNA live imaging. We are now excited to see the first applications of this new technique.
Disclosures
None declared
Wieland F and Béthune J: F1000Prime Recommendation of [Paige JS et al., Science 2011, 333(6042):642-6]. In F1000Prime, 04 Oct 2011; DOI: 10.3410/f.12351957.14692101. F1000Prime.com/12351957#eval14692101
10 Oct 2011
New Finding, Technical Advance
DOI: 10.3410/f.12351957.14704061
Spinach-tagged RNA is an outstanding tool that promises to develop the study of RNA intracellular dynamics in living cells at high temporal and spatial resolution. We are looking forward to seeing how it is applied in answering prominent questions in RNA biology.
Disclosures
None declared
Pontes O and Costa-Nunes P: F1000Prime Recommendation of [Paige JS et al., Science 2011, 333(6042):642-6]. In F1000Prime, 10 Oct 2011; DOI: 10.3410/f.12351957.14704061. F1000Prime.com/12351957#eval14704061
25 Oct 2011
Technical Advance
DOI: 10.3410/f.12351957.14726383
This is a very exciting advance in RNA imaging, providing the potential to study RNA dynamics in live cells with the ease (and multiplex options) with which protein dynamics have been studied for years using the available palette of fluorescent proteins.
Analysis of RNA dynamics has been complicated...Continue reading
Analysis of RNA dynamics has been complicated...Continue reading
This is a very exciting advance in RNA imaging, providing the potential to study RNA dynamics in live cells with the ease (and multiplex options) with which protein dynamics have been studied for years using the available palette of fluorescent proteins.
Analysis of RNA dynamics has been complicated by the necessity to co-express fluorescent proteins, which necessarily decrease the signal:noise ratio. Not only does this new approach solve this problem (the fluorophores are not fluorescent until they bind the RNA aptamer), but it already offers a range of fluorophores (which presumably could continue to expand) for multiplexed analysis of multiple RNAs. This significant technological advance is likely to have the same impact on the study of RNA dynamics as the discovery of the green fluorescent protein (GFP) has had on protein dynamics.
Analysis of RNA dynamics has been complicated by the necessity to co-express fluorescent proteins, which necessarily decrease the signal:noise ratio. Not only does this new approach solve this problem (the fluorophores are not fluorescent until they bind the RNA aptamer), but it already offers a range of fluorophores (which presumably could continue to expand) for multiplexed analysis of multiple RNAs. This significant technological advance is likely to have the same impact on the study of RNA dynamics as the discovery of the green fluorescent protein (GFP) has had on protein dynamics.
Disclosures
None declared
Trinkle-Mulcahy L: F1000Prime Recommendation of [Paige JS et al., Science 2011, 333(6042):642-6]. In F1000Prime, 25 Oct 2011; DOI: 10.3410/f.12351957.14726383. F1000Prime.com/12351957#eval14726383
07 Nov 2011
New Finding, Technical Advance
DOI: 10.3410/f.12351957.14730176
Although I fully endorse the other evaluations regarding the importance of following the fate of RNAs in live cells, for me the reason why this paper is really interesting is because it shows that, under the right conditions of selection, one can get RNA to carry out the function of a protein. This...Continue reading
Although I fully endorse the other evaluations regarding the importance of following the fate of RNAs in live cells, for me the reason why this paper is really interesting is because it shows that, under the right conditions of selection, one can get RNA to carry out the function of a protein. This is not only a promising development for biotechnology but it also strongly reinforces the idea that, at the origin of life, the first enzymes were almost certainly just folded up RNA molecules.
Disclosures
None declared
Joly E: F1000Prime Recommendation of [Paige JS et al., Science 2011, 333(6042):642-6]. In F1000Prime, 07 Nov 2011; DOI: 10.3410/f.12351957.14730176. F1000Prime.com/12351957#eval14730176
09 Dec 2011
Dissent
DOI: 10.3410/f.12351957.14763067
While this is very nice paper, I think the title and some of the comments are misleading.
GFP does not just bind a fluorescent compound, it produces the chromophore de novo from the protein itself, i.e. it is an enzyme that performs self-surgery.
The RNA described here mimics SOME aspects of GFP....Continue reading
GFP does not just bind a fluorescent compound, it produces the chromophore de novo from the protein itself, i.e. it is an enzyme that performs self-surgery.
The RNA described here mimics SOME aspects of GFP....Continue reading
While this is very nice paper, I think the title and some of the comments are misleading.
GFP does not just bind a fluorescent compound, it produces the chromophore de novo from the protein itself, i.e. it is an enzyme that performs self-surgery.
The RNA described here mimics SOME aspects of GFP. I completely agree that it will be a useful tool, but the selection process has another serious hurdle before the RNA has achieved the chemistry of this protein.
GFP does not just bind a fluorescent compound, it produces the chromophore de novo from the protein itself, i.e. it is an enzyme that performs self-surgery.
The RNA described here mimics SOME aspects of GFP. I completely agree that it will be a useful tool, but the selection process has another serious hurdle before the RNA has achieved the chemistry of this protein.
Disclosures
None declared
Phillips G: F1000Prime Dissenting Opinion on [Paige JS et al., Science 2011, 333(6042):642-6]. In F1000Prime, 09 Dec 2011; DOI: 10.3410/f.12351957.14763067. F1000Prime.com/12351957#eval14763067
F1000Prime Recommendations, Dissents and Comments for [Paige JS et al., Science 2011, 333(6042):642-6]. In F1000Prime, 22 May 2013; F1000Prime.com/12351957
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Green fluorescent protein (GFP) and its derivatives have transformed the use and analysis of proteins for diverse applications. Like proteins, RNA has complex roles in cellular function and is increasingly used for various applications, but a comparable approach for fluorescently tagging RNA is lacking. Here, we describe the generation of RNA aptamers that bind fluorophores resembling the fluorophore in GFP. These RNA-fluorophore complexes create a palette that spans the visible spectrum. An RNA-fluorophore complex, termed Spinach, resembles enhanced GFP and emits a green fluorescence comparable in brightness with fluorescent proteins. Spinach is markedly resistant to photobleaching, and Spinach fusion RNAs can be imaged in living cells. These RNA mimics of GFP provide an approach for genetic encoding of fluorescent RNAs.
PMID: 21798953
