Protein_Domain
NpuDnaE(C)

Part:BBa_K1362401:Experience

Designed by: Constantin Ahlmann-Eltze, Charlotte Bunne, Magdalena Büscher, Jan Gleixner, Max Horn, Anna Huhn, Nils Klughammer, Jakob Kreft, Elisabeth Schäfer, Carolin Schmelas, Silvan Schmitz, Max Waldha   Group: iGEM14_Heidelberg   (2014-10-06)


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Applications of BBa_K1362401


Characterization in Nicotiana benthamiana

Group: [http://2016.igem.org/Team:Valencia_UPV Valencia_UPV team 2016]

Author: Valencia UPV iGEM team (Iván Casas Rodrigo, Mónica Victoria Gutiérrez Salazar, Alicia Climent Catalá, Ãlvaro Ballesteros González)

Summary

Valencia UPV 2016 team tested and demonstrated the functionality of inteins in a plant chassis, specifically in Nicotiana benthamiana. We used the inteins with our split-Cas9 (BBa_K2017000 and BBa_K2017001). The inteins allowed to fusion of the two parts of the split-Cas9 in vivo, in order to obtain a fully functional Cas9 protein. In Figure 1 it is shown an electrophoresis gel with the results of the testing of split-Cas9 in plants. For further information about the experimental design, check the information of split-Cas9 parts, BBa_K2017000 and BBa_K2017001

Figure 1: Electrophoresis gel of XT1 amplicons digestion. Mutation efficiencies of Split-intein-Cas9 system and classical Cas9 system are compared by resistant bands signal intensity.


Additional information

  • Team:iGEM-IISERPUNE-2020
  • We aimed to use this part for circularization of the cyclotide protein. To build the biobrick we preferred restriction-free cloning but we realised that the biobrick was not compatible with restriction-free cloning. So we modified the part and built the modified version of C- Intein which is compatible with restriction-free cloning technique: BBa_K3582020.

User Reviews

UNIQa686545debd6a153-partinfo-00000000-QINU UNIQa686545debd6a153-partinfo-00000001-QINU