Difference between revisions of "Part:BBa K782067"
 
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* TALD label represents TAL effector 1295 from zebrafish experiments (Sander et al., 2011).
 
* TALD label represents TAL effector 1295 from zebrafish experiments (Sander et al., 2011).
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* Binding sites for individual TAL effectors are indicated with square brackets [ ].
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==Introduction==
 
==Introduction==
  
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Transcription activation like (TAL) effectors are bacterial plant pathogen transcription factors that bind to DNA by recognizing a specific DNA sequence in which each base pair binds a single tandem repeat in in the TAL DNA-binding domain. A tandem TAL repeat contains 33 to 35 amino acids, where the 12th and the 13th amino acid, called a “repeat variable diresidue” (RVD)  are responsible for specific interactions with the corresponding base pair (Scholze and Boch, 2011).  All TAL repeats have almost identical sequences, differing only in the RVDs. This modularity of TAL effector binding domains therefore makes them a perfect tool to target specific DNA sequences by designing specific binding domains for a selected TAL effector. We designed two consecutive specific binding sites for [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782011 NicTAL12:KRAB] and [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782009 TALD:KRAB](Figure 1).  
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Our construct contains two specific binding sites for [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782007 NicTAL12] and [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782005 TALD].  
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Single binding sequence for NicTAL is: TCTATCAATGATAGA
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Single binding sequence for NicTAL12 is: TCTATCAATGATAGA  
  
 
Single binding sequence for TALD is: TCGTCCAATAGCTTCTC
 
Single binding sequence for TALD is: TCGTCCAATAGCTTCTC
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[[Image:2xNic+2xD.png‎]]
 
[[Image:2xNic+2xD.png‎]]
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'''Figure 1.''' Shematic representation of two consecutive specific binding sites for NicTAL and TALD.
 
  
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'''Figure 1.''' Shematic representation of two consecutive specific binding sites for NicTAL12 and TALD.
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* Binding sites for TAL effectors were ordered from GeneArt.
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==Characterization==
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Specific TAL binding sites were further characterized with different reporter constructs.
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'''Results:'''
  
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* [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782024 BBa_K782024]
  
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* Binding sites for TAL effectors were ordered from IDT.
 
  
  
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Scholze, H., and Boch, J. (2011) TAL effectors are remote controls for gene activation. Curr. Opin. Microbiol. 14, 47-53.
 
Scholze, H., and Boch, J. (2011) TAL effectors are remote controls for gene activation. Curr. Opin. Microbiol. 14, 47-53.
  
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Sander, J. D., Cade, L., Khayter, C., Reyon, D., Peterson, R. T., Joung, J. K., and Yeh, J.-R. J. (2011) Targeted gene disruption in somatic zebrafish cells using engineered TALENs. Nature Biotechnology 29, 697–698  
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Sander, J. D., Cade, L., Khayter, C., Reyon, D., Peterson, R. T., Joung, J. K., and Yeh, J.-R. J. (2011) Targeted gene disruption in somatic zebrafish cells using engineered TALENs. Nature Biotechnology 29, 697–698.
  
  

Latest revision as of 22:50, 26 September 2012

2x[NicTAL]+2x[TALD] operator

  • TALD label represents TAL effector 1295 from zebrafish experiments (Sander et al., 2011).
  • Binding sites for individual TAL effectors are indicated with square brackets [ ].


Introduction

Our construct contains two specific binding sites for NicTAL12 and TALD.

Single binding sequence for NicTAL12 is: TCTATCAATGATAGA

Single binding sequence for TALD is: TCGTCCAATAGCTTCTC


2xNic+2xD.png

Figure 1. Shematic representation of two consecutive specific binding sites for NicTAL12 and TALD.


  • Binding sites for TAL effectors were ordered from GeneArt.


Characterization

Specific TAL binding sites were further characterized with different reporter constructs.

Results:


References

Scholze, H., and Boch, J. (2011) TAL effectors are remote controls for gene activation. Curr. Opin. Microbiol. 14, 47-53.

Sander, J. D., Cade, L., Khayter, C., Reyon, D., Peterson, R. T., Joung, J. K., and Yeh, J.-R. J. (2011) Targeted gene disruption in somatic zebrafish cells using engineered TALENs. Nature Biotechnology 29, 697–698.


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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 35
    Illegal AgeI site found at 105
  • 1000
    COMPATIBLE WITH RFC[1000]