Difference between revisions of "Part:BBa K2549021"
m
Line 3: Line 3:
 
<partinfo>BBa_K2549021 short</partinfo>
 
<partinfo>BBa_K2549021 short</partinfo>
  
−
This part is one of our SynNotch receptors<ref>Engineering Customized Cell Sensing and Response Behaviors Using Synthetic Notch Receptors. Morsut L, Roybal KT, Xiong X, ..., Thomson M, Lim WA. Cell, 2016 Feb;164(4):780-91 PMID: 26830878; DOI: 10.1016/j.cell.2016.01.012</ref>. ACD19 ([[Part:BBa_K2549005]]) is used as the extracellular sensor module to receive the signal input from surface-expressed CD19. mN1c ([[Part:BBa_K2549006]]) is served as the transmembrane core domain of SynNotch, which is evident to have a low basal expression and a high activation efficiency. tTAA ([[Part:BBa_K2446057]]) is an improved tetracycline-controlled transactivator, which is cleaved after SynNotch activation and drives the expression of the amplifier. Besides, a CD8&alpha; signal peptide ([[Part:BBa_K2549044]]) and a Myc-tag ([[Part:BBa_K823036]]) are added to the N terminal of ACD19 ([[Part:BBa_K2549005]]) for membrane targeting and easy determination of surface expression.   
+
This part is one of our SynNotch receptors, similar to the original published version<ref>Engineering Customized Cell Sensing and Response Behaviors Using Synthetic Notch Receptors. Morsut L, Roybal KT, Xiong X, ..., Thomson M, Lim WA. Cell, 2016 Feb;164(4):780-91 PMID: 26830878; DOI: 10.1016/j.cell.2016.01.012</ref>. It is our favorite. &alpha;CD19 ([[Part:BBa_K2549005]]) is used as the extracellular sensor module to receive the signal input from surface-expressed CD19. mN1c ([[Part:BBa_K2549006]]) is served as the transmembrane core domain of SynNotch, which is evident to have a low basal expression and a high activation efficiency. tTAA ([[Part:BBa_K2446057]]) is an improved tetracycline-controlled transactivator, which is cleaved after SynNotch activation and drives the expression of the amplifier. Besides, a CD8&alpha; signal peptide ([[Part:BBa_K2549044]]) and a Myc-tag ([[Part:BBa_K823036]]) are added to the N terminal of &alpha;CD19 ([[Part:BBa_K2549005]]) for membrane targeting and easy determination of surface expression.   
 +
 
 +
<!-- -->
 +
<span class='h3bb'>Sequence and Features</span>
 +
<partinfo>BBa_K2549021 SequenceAndFeatures</partinfo>
 +
 
  
 
<!-- Add more about the biology of this part here -->
 
<!-- Add more about the biology of this part here -->
 
===Biology===
 
===Biology===
−
====SynNotch receptors function well in Lim WA et al in 2016====
+
=====It works as expected=====
 +
@@@@
  
−
[[File:SynNotch.jpeg|none|360px|thumb|Lim WA et al stated:''SynNotch receptors provide extraordinary flexibility in engineering cells with customized sensing/response behaviors to user-specified extracellular cues.'']]
+
=====SynNotch receptors function well in Morsut L et al 2016=====
  
−
[[File:SynNotchECDandICD.jpeg|none|360px|thumb|Lim WA et al have shown that modularity of the synNotch platform. They stated : ''the input and output domains from Notch can be swapped with diverse domains. On the extracellular side, diverse recognition domains can be used (antibody based, or peptide tags are shown) and on the intracellular side, diverse effector can be used (transcriptional activators with different DNA-binding domains are shown, as well as a transcriptional repressor).'']]
+
[[File:SynNotch.jpeg|none|300px|thumb|Morsut L et al stated:''SynNotch receptors provide extraordinary flexibility in engineering cells with customized sensing/response behaviors to user-specified extracellular cues.'']]
  
−
Please refer to the original article for more details<ref>Engineering Customized Cell Sensing and Response Behaviors Using Synthetic Notch Receptors. Morsut L, Roybal KT, Xiong X, ..., Thomson M, Lim WA. Cell, 2016 Feb;164(4):780-91 PMID: 26830878; DOI: 10.1016/j.cell.2016.01.012</ref>.
+
[[File:SynNotchECDandICD.jpeg|none|400px|thumb|Morsut L et al have shown that modularity of the synNotch platform. They stated: ''the input and output domains from Notch can be swapped with diverse domains. On the extracellular side, diverse recognition domains can be used (antibody based, or peptide tags are shown) and on the intracellular side, diverse effector can be used (transcriptional activators with different DNA-binding domains are shown, as well as a transcriptional repressor).'']]
  
−
====ACD19-mN1c-tTAA works as expected====
+
Please refer to the original article for more details.
−
<!-- -->
+
−
<span class='h3bb'>Sequence and Features</span>
+
−
<partinfo>BBa_K2549021 SequenceAndFeatures</partinfo>
+
  
  

Revision as of 15:17, 12 October 2018


aCD19-mN1c-tTAA

This part is one of our SynNotch receptors, similar to the original published version[1]. It is our favorite. αCD19 (Part:BBa_K2549005) is used as the extracellular sensor module to receive the signal input from surface-expressed CD19. mN1c (Part:BBa_K2549006) is served as the transmembrane core domain of SynNotch, which is evident to have a low basal expression and a high activation efficiency. tTAA (Part:BBa_K2446057) is an improved tetracycline-controlled transactivator, which is cleaved after SynNotch activation and drives the expression of the amplifier. Besides, a CD8α signal peptide (Part:BBa_K2549044) and a Myc-tag (Part:BBa_K823036) are added to the N terminal of αCD19 (Part:BBa_K2549005) for membrane targeting and easy determination of surface expression.

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 NgoMIV site found at 2493
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 534
    Illegal SapI.rc site found at 1408


Biology

It works as expected

@@@@

SynNotch receptors function well in Morsut L et al 2016
Morsut L et al stated:SynNotch receptors provide extraordinary flexibility in engineering cells with customized sensing/response behaviors to user-specified extracellular cues.
Morsut L et al have shown that modularity of the synNotch platform. They stated: the input and output domains from Notch can be swapped with diverse domains. On the extracellular side, diverse recognition domains can be used (antibody based, or peptide tags are shown) and on the intracellular side, diverse effector can be used (transcriptional activators with different DNA-binding domains are shown, as well as a transcriptional repressor).

Please refer to the original article for more details.


References

  1. ↑ Engineering Customized Cell Sensing and Response Behaviors Using Synthetic Notch Receptors. Morsut L, Roybal KT, Xiong X, ..., Thomson M, Lim WA. Cell, 2016 Feb;164(4):780-91 PMID: 26830878; DOI: 10.1016/j.cell.2016.01.012