Difference between revisions of "Part:BBa K2549032"
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<partinfo>BBa_K2549032 short</partinfo> | <partinfo>BBa_K2549032 short</partinfo> | ||
| − | This part is one of the response elements of our amplifier, also executing the combiner function. 4*ZF21.16 binding sites ([[Part:BBa_K2446008]]) and 2*ZF43.8 binding sites ([[Part:BBa_K2549051]]) can bind to different zinc finger-based transcription activator ZF21.16-VP64 ([[Part:BBa_K2549023]]) and zinc finger-based repressor ZF43.8-KRAB ([[Part:BBa_K2446041]]), respectively, with high orthogonality. Minimal CMV ([[Part:BBa_K2549049]]) is a promotor providing very low basal expression and high maximal expression after induction. This part | + | This part is one of the response elements of our amplifier, also executing the combiner function. 4*ZF21.16 binding sites ([[Part:BBa_K2446008]]) and 2*ZF43.8 binding sites ([[Part:BBa_K2549051]]) can bind to different zinc finger-based transcription activator ZF21.16-VP64 ([[Part:BBa_K2549023]]) and zinc finger-based repressor ZF43.8-KRAB ([[Part:BBa_K2446041]]), respectively, with high orthogonality. Minimal CMV ([[Part:BBa_K2549049]]) is a promotor providing very low basal expression and high maximal expression after induction. This part was designed to construct our NIMPLY logic gate and test our multiple binding sites amplifier model<ref>http://2018.igem.org/Team:Fudan/Model#Transcriptional_Amplifer</ref>. |
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| + | <!-- --> | ||
| + | <span class='h3bb'>Sequence and Features</span> | ||
| + | <partinfo>BBa_K2549032 SequenceAndFeatures</partinfo> | ||
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<!-- Add more about the biology of this part here --> | <!-- Add more about the biology of this part here --> | ||
===Biology=== | ===Biology=== | ||
| − | ==== | + | =====It works as we designed ===== |
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| − | + | Different transcription factors interaction with multiple binding sites | |
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| − | + | Promotor operator numbers can have an impact on tuning output | |
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| − | + | =====synthetic promotor operators regulated by artificial zinc finger-based transcription factors===== | |
| − | + | Khalil AS et al have reported several synthetic promotor operators which can interact with artificial zinc finger-based transcription factors with high specificity and high orthogonality<ref>A synthetic biology framework for programming eukaryotic transcription functions. Khalil AS, Lu TK, Bashor CJ, ..., Joung JK, Collins JJ. Cell, 2012 Aug;150(3):647-58 PMID: 22863014; DOI: 10.1016/j.cell.2012.05.045</ref>. | |
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| + | [[File:ZF2.jpg|none|540px|thumb|Khalil AS et al stated:''sTFs constructed from OPEN-engineered ZFs are orthogonal to one another. sTF43-8 activated noncognate Promoter21-16 due to the fortuitous creation of a sequence that is significantly similar to the binding sequence of 43-8, when the downstream BamHI restriction site is considered.'']] | ||
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| + | [[File:ZF3.jpg|none|540px|thumb|Khalil AS et al stated:''Tuning up output strength by increasing ZF operator number in synthetic promoter (sTF43-8).'']] | ||
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===Functional Parameters=== | ===Functional Parameters=== | ||
<partinfo>BBa_K2549032 parameters</partinfo> | <partinfo>BBa_K2549032 parameters</partinfo> | ||
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===References=== | ===References=== | ||
Revision as of 14:07, 12 October 2018
4*ZF21.16-minCMV-2*ZF43.8
This part is one of the response elements of our amplifier, also executing the combiner function. 4*ZF21.16 binding sites (Part:BBa_K2446008) and 2*ZF43.8 binding sites (Part:BBa_K2549051) can bind to different zinc finger-based transcription activator ZF21.16-VP64 (Part:BBa_K2549023) and zinc finger-based repressor ZF43.8-KRAB (Part:BBa_K2446041), respectively, with high orthogonality. Minimal CMV (Part:BBa_K2549049) is a promotor providing very low basal expression and high maximal expression after induction. This part was designed to construct our NIMPLY logic gate and test our multiple binding sites amplifier model[1].
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Biology
It works as we designed
Different transcription factors interaction with multiple binding sites
Promotor operator numbers can have an impact on tuning output
synthetic promotor operators regulated by artificial zinc finger-based transcription factors
Khalil AS et al have reported several synthetic promotor operators which can interact with artificial zinc finger-based transcription factors with high specificity and high orthogonality[2].
Khalil AS et al stated:sTFs constructed from OPEN-engineered ZFs are orthogonal to one another. sTF43-8 activated noncognate Promoter21-16 due to the fortuitous creation of a sequence that is significantly similar to the binding sequence of 43-8, when the downstream BamHI restriction site is considered.
