Part:BBa_K1362400:Experience

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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, Álvaro 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 N-Intein which is compatible with restriction-free cloning technique: BBa_K3582022.

User Reviews

UNIQf32cd96b2277db1f-partinfo-00000000-QINU UNIQf32cd96b2277db1f-partinfo-00000001-QINU