Difference between revisions of "Part:BBa K5490030"

 
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PLoS One. 2017 Feb 27;12(2):e0171257. doi: 10.1371/journal.pone.0171257. PMID: 28241009; PMCID: PMC5328254.IRES sequences are typically derived from viruses and can be inserted between two open reading frames (ORFs) in a single mRNA. This strategic placement allows for the translation of two proteins from the same polycistronic mRNA. IRES elements facilitate ribosome recruitment to the downstream ORF, enabling independent initiation of translation without requiring a 5' cap structure.
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IRES sequences are widely applied in molecular biology, most notably as ribosome binding sites (RBS) in bicistronic or multicistronic constructs. They are especially useful when co-expressing multiple genes or proteins, such as in gene therapy, vaccine development, or studies requiring simultaneous expression of reporter and target genes.
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Fitzgerald KD, Semler BL. Bridging IRES elements in mRNAs to the eukaryotic translation apparatus. Biochim Biophys Acta. 2009 Sep-Oct;1789(9-10):518-28. doi: 10.1016/j.bbagrm.2009.07.004. Epub 2009 Jul 23. PMID: 19631772; PMCID: PMC2783899.
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mCherry is one of the most popular fluorescent reporters, belonging to the DsRed family of fluorescent proteins. Its red color is particularly useful for multicolor monitoring of different target genes in live-cell imaging, as it minimizes overlap with green and blue fluorescent proteins like GFP or CFP.
 +
 +
Compared to earlier variants of red fluorescent proteins, mCherry offers several key improvements. Notably, it is monomeric, making it ideal for fusing to other proteins without disrupting their function or causing aggregation. This feature enhances its utility in protein tracking and functional studies.
 +
 +
mCherry is also non-toxic, enabling its use in live-cell applications without compromising cell viability. Furthermore, it retains fluorescence across a broad range of pH levels, making it suitable for imaging in various intracellular environments.
 +
 +
Another advantage is its stability and resistance to photobleaching, allowing for long-term monitoring of samples under the microscope. These qualities make mCherry a reliable tool for dynamic imaging in fields like developmental biology, neuroscience, and cancer research.
 +
 +
Huttly A. Reporter genes. Methods Mol Biol. 2009;478:39-69. doi: 10.1007/978-1-59745-379-0_3. PMID: 19009438.
 +
Shen Y, Chen Y, Wu J, Shaner NC, Campbell RE. Engineering of mCherry variants with long Stokes shift, red-shifted fluorescence, and low cytotoxicity.

Revision as of 00:33, 29 September 2024


IRES2

Viral RBS for mCherryNLS

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


PLoS One. 2017 Feb 27;12(2):e0171257. doi: 10.1371/journal.pone.0171257. PMID: 28241009; PMCID: PMC5328254.IRES sequences are typically derived from viruses and can be inserted between two open reading frames (ORFs) in a single mRNA. This strategic placement allows for the translation of two proteins from the same polycistronic mRNA. IRES elements facilitate ribosome recruitment to the downstream ORF, enabling independent initiation of translation without requiring a 5' cap structure.

IRES sequences are widely applied in molecular biology, most notably as ribosome binding sites (RBS) in bicistronic or multicistronic constructs. They are especially useful when co-expressing multiple genes or proteins, such as in gene therapy, vaccine development, or studies requiring simultaneous expression of reporter and target genes. Fitzgerald KD, Semler BL. Bridging IRES elements in mRNAs to the eukaryotic translation apparatus. Biochim Biophys Acta. 2009 Sep-Oct;1789(9-10):518-28. doi: 10.1016/j.bbagrm.2009.07.004. Epub 2009 Jul 23. PMID: 19631772; PMCID: PMC2783899.

mCherry is one of the most popular fluorescent reporters, belonging to the DsRed family of fluorescent proteins. Its red color is particularly useful for multicolor monitoring of different target genes in live-cell imaging, as it minimizes overlap with green and blue fluorescent proteins like GFP or CFP.

Compared to earlier variants of red fluorescent proteins, mCherry offers several key improvements. Notably, it is monomeric, making it ideal for fusing to other proteins without disrupting their function or causing aggregation. This feature enhances its utility in protein tracking and functional studies.

mCherry is also non-toxic, enabling its use in live-cell applications without compromising cell viability. Furthermore, it retains fluorescence across a broad range of pH levels, making it suitable for imaging in various intracellular environments.

Another advantage is its stability and resistance to photobleaching, allowing for long-term monitoring of samples under the microscope. These qualities make mCherry a reliable tool for dynamic imaging in fields like developmental biology, neuroscience, and cancer research.

Huttly A. Reporter genes. Methods Mol Biol. 2009;478:39-69. doi: 10.1007/978-1-59745-379-0_3. PMID: 19009438. Shen Y, Chen Y, Wu J, Shaner NC, Campbell RE. Engineering of mCherry variants with long Stokes shift, red-shifted fluorescence, and low cytotoxicity.