Difference between revisions of "Part:BBa K782072"
 
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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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* DNA 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 twelve specific binding sites for TALD (Figure 1).  
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TAL effectors (TALEs) are able to specifically bind to the DNA by recognizing base pairs on a DNA molecule. With TAL regulators we are able to control expression of a desired gene. Efficient transcription control can be achieved by binding of multiple TAL regulators upstream of promoter.
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Our construct contain twelve specific binding sites for TALD (Figure 1).  
  
 
Single binding sequence for [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782005 TALD] is: TCGTCCAATAGCTTCTC
 
Single binding sequence for [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782005 TALD] is: TCGTCCAATAGCTTCTC
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[[Image:12xD.png]]
 
[[Image:12xD.png]]
  
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'''Figure 1.''' Shematic representation of twelve specific binding site for TALD
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'''Figure 1.''' Shematic representation of the construct.
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upstream of CMV promoter and reporter protein mCitrine.  
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* mCitrine was provided from host lab.
 
 
* Binding sites for TAL effectors were ordered from IDT.  
 
* Binding sites for TAL effectors were ordered from IDT.  
  
  
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==References==
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==Characterization==  
  
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Scholze, H., and Boch, J. (2011) TAL effectors are remote controls for gene activation. Curr. Opin. Microbiol. 14, 47-53.
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Construct was later used for constructing different reporter plasmids to show specific binding of [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782009 TALD:KRAB]/[https://parts.igem.org/wiki/index.php?title=Part:BBa_K782012 TALD:VP16].
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'''Results:'''
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* [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782031 BBa_K782031]
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* [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782022 BBa_K782022]
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==References==
  
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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 22:55, 26 September 2012

12x[TALD] operator

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


Introduction

TAL effectors (TALEs) are able to specifically bind to the DNA by recognizing base pairs on a DNA molecule. With TAL regulators we are able to control expression of a desired gene. Efficient transcription control can be achieved by binding of multiple TAL regulators upstream of promoter.

Our construct contain twelve specific binding sites for TALD (Figure 1).

Single binding sequence for TALD is: TCGTCCAATAGCTTCTC


12xD.png

Figure 1. Shematic representation of the construct.


  • Binding sites for TAL effectors were ordered from IDT.


Characterization

Construct was later used for constructing different reporter plasmids to show specific binding of TALD:KRAB/TALD:VP16.

Results:


References

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 214
    Illegal AgeI site found at 446
  • 1000
    COMPATIBLE WITH RFC[1000]