Difference between revisions of "Part:BBa K5490001"

 
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gRNAs (guide RNAs) are relatively small RNA molecules that play a crucial role in gene editing and RNA-targeting technologies. They are typically expressed under the control of a promoter, often polymerase III, which is ideal given the high complexity of the gRNA's secondary structure. These gRNAs are composed of two main components:
 
gRNAs (guide RNAs) are relatively small RNA molecules that play a crucial role in gene editing and RNA-targeting technologies. They are typically expressed under the control of a promoter, often polymerase III, which is ideal given the high complexity of the gRNA's secondary structure. These gRNAs are composed of two main components:
  
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The scaffold or direct repeat region: This region forms a complex secondary structure after transcription, allowing it to bind effectively to an effector molecule such as CasRx, which acts as an RNA nuclease, specifically targeting single-stranded RNA. There are two types of direct repeat sequences commonly used--one that is 30 nucleotides long and another that is 36 nucleotides long. The 36-nucleotide direct repeat has been shown to have a higher affinity for CasRx, improving the overall efficiency of the RNA-targeting system.
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The scaffold or direct repeat region
  
 
The spacer: This is a 23-nucleotide sequence that is complementary to the target RNA, guiding CasRx to the specific RNA region for cleavage. In this case, the target is a region within the West Nile Virus (WNV) genome. The spacer sequence was selected using an algorithm designed to optimize its binding and cutting efficiency, ensuring that the CasRx-gRNA complex can cleave the target RNA with high specificity and effectiveness.
 
The spacer: This is a 23-nucleotide sequence that is complementary to the target RNA, guiding CasRx to the specific RNA region for cleavage. In this case, the target is a region within the West Nile Virus (WNV) genome. The spacer sequence was selected using an algorithm designed to optimize its binding and cutting efficiency, ensuring that the CasRx-gRNA complex can cleave the target RNA with high specificity and effectiveness.
  
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By integrating both the scaffold and spacer, researchers can achieve precise RNA cleavage. The CasRx effector dimer, guided by the gRNA, binds to the target RNA region, enabling highly efficient RNA degradation. This optimized approach allows for better control and precision in RNA-targeting applications, such as combating viral infections like WNV or studying gene expression pathways.
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By integrating both the scaffold and spacer, researchers can achieve precise RNA cleavage. This is the second spacer sequence.
  
  

Revision as of 16:23, 24 September 2024


23-nt sequence binds CasRx to cleave WNV genome; modifiable target 2

gRNAs (guide RNAs) are relatively small RNA molecules that play a crucial role in gene editing and RNA-targeting technologies. They are typically expressed under the control of a promoter, often polymerase III, which is ideal given the high complexity of the gRNA's secondary structure. These gRNAs are composed of two main components:

The scaffold or direct repeat region

The spacer: This is a 23-nucleotide sequence that is complementary to the target RNA, guiding CasRx to the specific RNA region for cleavage. In this case, the target is a region within the West Nile Virus (WNV) genome. The spacer sequence was selected using an algorithm designed to optimize its binding and cutting efficiency, ensuring that the CasRx-gRNA complex can cleave the target RNA with high specificity and effectiveness.

By integrating both the scaffold and spacer, researchers can achieve precise RNA cleavage. This is the second spacer sequence.


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]