Part:BBa_K5490025

mCherry NLS

An mCherry fluorescent protein is separately expressed and directed to the nucleus via a nuclear localization signal (NLS). The purpose of mCherry is to act as a reporter, enabling independent monitoring of the CasRx activity and target RNA levels. This separation of signals allows for precise tracking of the effector molecule (CasRx) and the concentration of the target RNA, enhancing the accuracy of the experiment.

By monitoring mCherry fluorescence in the nucleus, researchers can confirm the presence of CasRx without interference from the target RNA cleavage activity, ensuring clear and accurate results in gene silencing studies or antiviral applications.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 352
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 352
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 352
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 352
  
• 1000
  COMPATIBLE WITH RFC[1000]

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.