Part:BBa_K5490028

CasRx It’s an inducible endonuclease protein that can target any RNA sequence complementary to a gRNA. It contains an HA tag for Western blot and Immunohistochemistry analysis, and a mchery reporter with an NLS for separating activity between CasRx and the target RNA.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 499
  Illegal EcoRI site found at 919
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 499
  Illegal EcoRI site found at 919
  Illegal NotI site found at 2908
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 499
  Illegal EcoRI site found at 919
  Illegal BamHI site found at 2899
  Illegal XhoI site found at 2374
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 499
  Illegal EcoRI site found at 919
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 499
  Illegal EcoRI site found at 919
  Illegal NgoMIV site found at 187
  
• 1000
  COMPATIBLE WITH RFC[1000]

CasRx is a highly effective RNA-targeting molecule with nuclease activity specifically against single-stranded RNA, making it a versatile tool for gene silencing and antiviral therapies. One of its major advantages over RNA interference (RNAi) is its lower off-target activity, providing more preecision in targeting specific RNA sequences. As a viral protein, CasRx operates independently of the host's cellular machinery, which reduces variability and enhances efficiency.

CasRx is the smallest member of the RNA-targeting CRISPR family, allowing for easier delivery into cells while maintaining high specificity and efficiency in cleaving target RNA. Its minimal off-target effects and high efficiency make it ideal for large-scale transcriptome screening, precise gene silencing, and targeting viral RNA in therapeutic applications.

A key advantage of CasRx over other CRISPR systems is its simplicity--it does not require a PAM sequence or a tracrRNA, making it one of the most straightforward CRISPR systems to implement. CasRx can target virtually any RNA sequence when paired with a guide RNA (gRNA) that is complementary to the target.

Konermann S, Lotfy P, Brideau NJ, Oki J, Shokhirev MN, Hsu PD. Transcriptome Engineering with RNA-Targeting Type VI-D CRISPR Effectors. Cell. 2018 Apr 19;173(3):665-676.e14. doi: 10.1016/j.cell.2018.02.033. Epub 2018 Mar 15. PMID: 29551272; PMCID: PMC5910255. Chuang YF, Wang PY, Kumar S, Lama S, Lin FL, Liu GS. Methods for in vitro CRISPR/CasRx-Mediated RNA Editing. Front Cell Dev Biol. 2021 Jun 11;9:667879. doi: 10.3389/fcell.2021.667879. PMID: 34178991; PMCID: PMC8226256.