Difference between revisions of "Part:BBa K2200003"
 
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==Contribution by iGEM22 Team CUHK-Hong Kong-SBS==
  
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Atreus testing
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​​U6 small nuclear promoter is one of the promoters that are used to drive the expression of small RNAs (Gao, Herrera-Carrillo & Berkhout, 2018). U6 promoter is a Type 3 RNA polymerase III promoters that are responsible for the transcription of small RNAs in eukaryotes(ibid). As a Type 3 promoter, it is superior for small RNA expression cassettes for plasmid design and small RNA delivery in therapy because of their simple structure, well-studied transcription start site(ibid).U6 promoter is also unique among the three types of RNA polymerase 3 promoters given that they only requires upstream regulatory elements which resemble the mRNA-type Polymerase 3 promoters(ibid).
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For the structure of U6 promoter, it is composed of characteristic promoter elements such as the enhancer and core regions (Ma & Hernandez, 2002). The core regions include a proximal sequence element (PSE) and a TATA box located about 50 and 25 bp respectively, upstream of the transcription start site(ibid). The PSE would be required for recruitment of snRNA-activating protein complex (SNAPc) and the TATA box would be necessary for binding to the TATA box binding protein for basal transcription(ibid).
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Regarding its efficiency, in a study of the Type 3 RNA polymerase 3 promoters, it is shown that all the 7SK, U6, and H1 promoters exhibit similar Polymerase 3 activity and successfully drive the transcription of luciferase (Gao, Herrera-Carrillo & Berkhout, 2018). In another literature, it has also been suggested that the efficiencies of U6 promoter is higher than that of H1 promoter for transcription of GFP small silencing RNA (siRNA) with lentiviral vectors in in vitro study and mouse brain (Mäkinen, 2006).
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'''Reference'''
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Gao Z.L., Herrera-Carrillo, E., & Berkhout, B. (2018). RNA Polymerase II Activity of Type 3 Pol III Promoters. Molecular Therapy. Nucleic Acids, 12, 135–145. https://doi.org/10.1016/j.omtn.2018.05.001
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Ma, B& Hernandez, N. (2002). Redundant Cooperative Interactions for Assembly of a Human U6 Transcription Initiation Complex. Molecular and Cellular Biology, 22(22), 8067–8078. https://doi.org/10.1128/MCB.22.22.8067-8078.2002
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Mäkinen, P. I., Koponen, J. K., Kärkkäinen, A.-M., Malm, T. M., Pulkkinen, K. H., Koistinaho, J., Turunen, M. P., & Ylä-Herttuala, S. (2006). Stable RNA interference: comparison of U6 and H1 promoters in endothelial cells and in mouse brain. The Journal of Gene Medicine, 8(4), 433–441. https://doi.org/10.1002/jgm.860

Latest revision as of 05:41, 30 September 2022


Human U6 promoter is a common part for eukaryotic expression systems.

Human U6 promoter is a common part for eukaryotic expression systems. We use it to initiate the transcription of sgRNA.

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]


Contribution by iGEM22 Team CUHK-Hong Kong-SBS

​​U6 small nuclear promoter is one of the promoters that are used to drive the expression of small RNAs (Gao, Herrera-Carrillo & Berkhout, 2018). U6 promoter is a Type 3 RNA polymerase III promoters that are responsible for the transcription of small RNAs in eukaryotes(ibid). As a Type 3 promoter, it is superior for small RNA expression cassettes for plasmid design and small RNA delivery in therapy because of their simple structure, well-studied transcription start site(ibid).U6 promoter is also unique among the three types of RNA polymerase 3 promoters given that they only requires upstream regulatory elements which resemble the mRNA-type Polymerase 3 promoters(ibid).

For the structure of U6 promoter, it is composed of characteristic promoter elements such as the enhancer and core regions (Ma & Hernandez, 2002). The core regions include a proximal sequence element (PSE) and a TATA box located about 50 and 25 bp respectively, upstream of the transcription start site(ibid). The PSE would be required for recruitment of snRNA-activating protein complex (SNAPc) and the TATA box would be necessary for binding to the TATA box binding protein for basal transcription(ibid).

Regarding its efficiency, in a study of the Type 3 RNA polymerase 3 promoters, it is shown that all the 7SK, U6, and H1 promoters exhibit similar Polymerase 3 activity and successfully drive the transcription of luciferase (Gao, Herrera-Carrillo & Berkhout, 2018). In another literature, it has also been suggested that the efficiencies of U6 promoter is higher than that of H1 promoter for transcription of GFP small silencing RNA (siRNA) with lentiviral vectors in in vitro study and mouse brain (Mäkinen, 2006).

Reference

Gao Z.L., Herrera-Carrillo, E., & Berkhout, B. (2018). RNA Polymerase II Activity of Type 3 Pol III Promoters. Molecular Therapy. Nucleic Acids, 12, 135–145. https://doi.org/10.1016/j.omtn.2018.05.001

Ma, B& Hernandez, N. (2002). Redundant Cooperative Interactions for Assembly of a Human U6 Transcription Initiation Complex. Molecular and Cellular Biology, 22(22), 8067–8078. https://doi.org/10.1128/MCB.22.22.8067-8078.2002

Mäkinen, P. I., Koponen, J. K., Kärkkäinen, A.-M., Malm, T. M., Pulkkinen, K. H., Koistinaho, J., Turunen, M. P., & Ylä-Herttuala, S. (2006). Stable RNA interference: comparison of U6 and H1 promoters in endothelial cells and in mouse brain. The Journal of Gene Medicine, 8(4), 433–441. https://doi.org/10.1002/jgm.860