Our team has created some new parts, mainly including the construction of chromosomal stabilization element group, tags, and some parts used in the construction of single base editor.
Chromosomal stabilization element group:
In order to transform centromeres, the core part of our project, into the genome of Saccharomyces cerevisiae cells, we constructed several fragments for transformation. In order to ensure the validity of transformation, we added tags for screening and corresponding promoters and terminators.
>>Please refer to our result page for more details.
Chromosome transfer (mRFP):
In order to directly observe the results of the chromosome transfer after fusion, we inducted mRFP into Saccharomyces cerevisiae, so that we can take photos of the fusants with fluorescence microscope. As a result, we created the following parts and proved them to work properly.
>>Please refer to our result page for more details.
Single base editor section:
We have created a fusion characterization system using CRISPR-dcas9 technology, which is mainly composed of mutant dCas9 fusion protein, gRNA and protospacer. The fusion protein consists of adenine base editor (ABE) and dcas9 protein, which are linked by a rigid linker, and its expression is controlled by PGK1 promoter and terminator. Through its combination with gRNA, we can see that it performs a certain degree of timing editing on the corresponding protospacer. This Biobrick can be used not only for the fusion characterization of these two cells, but also for the recording of specific events in certain cells after iGEM.
>>Please refer to our result page for more details.
Parts Table
Basic Parts
Name | Type | Description |
---|---|---|
BBa_K3112000 | DNA | S.C cenXV |
BBa_K3112001 | DNA | Y.L cenE |
BBa_K3112002 | Coding | dCas9 |
BBa_K3112004 | Regulatory | TEF promoter |
BBa_K3112005 | Terminator | CYC1 Terminator |
BBa_K3112007 | RNA | gRNA |
BBa_K3112008 | DNA | Protospacer+PAM |
BBa_K3112009 | Coding | GFP |
BBa_K3112010 | Coding | mRFP use in yeast |
BBa_K3112011 | Regulatory | PGK1 Promoter |
BBa_K3112012 | Terminator | PGK1 Terminator |
BBa_K3112013 | Coding | yEGFP |
BBa_K3112014 | Regulatory | His3 Promoter |
BBa_K3112015 | Coding | His3 Gene |
BBa_K3112016 | Regulatory | LEU2 Promoter |
BBa_K3112017 | Coding | LEU2 Gene |
BBa_K3112018 | Regulatory | URA3 Promoter |
BBa_K3112019 | Coding | URA3 Gene |
BBa_K3112020 | Coding | Hygromycin Resistance Gene(hyg or hph) |
BBa_K3112021 | Coding | A to G adenine Base Editor |
BBa_K3112022 | Terminator | TEF Terminator |
BBa_K3112023 | Coding | APOBEC1 cytidine deaminase |
BBa_K3112024 | Regulatory | GAP Promoter |
BBa_K3112025 | Terminator | ADH1 Terminator |
BBa_K3112026 | DNA | Homologous arm in Yarrowia lipolytica E Chromosome 1 |
BBa_K3112027 | DNA | Homologous arm in Yarrowia lipolytica E Chromosome 2 |
BBa_K3112028 | DNA | Homologous arm in Saccharomyces cerevisiae V Chromosome 1 |
BBa_K3112029 | DNA | Homologous arm in Saccharomyces cerevisiae V Chromosome 2 |
BBa_K3112030 | DNA | Homologous arm in Saccharomyces cerevisiae V Chromosome 3 |
BBa_K3112031 | DNA | Homologous arm in Saccharomyces cerevisiae V Chromosome 4 |
BBa_K3112032 | DNA | Left Homologous Arm for Homologous Recombination of Saccharomyces cerevisiae |
BBa_K3112033 | DNA | Right Homologous Arm for Homologous Recombination of Saccharomyces cerevisiae |
BBa_K3112034 | DNA | Left Homologous Arm for Homologous Recombination of Yarrowia lipolytica |
BBa_K3112035 | DNA | Right Homologous Arm for Homologous Recombination of Yarrowia lipolytica |
BBa_K3112037 | Coding | KanMX |
Composite Parts
Name | Type | Description |
---|---|---|
BBa_K3112040 | Composite | Chromosome stabilizing elements example 1 |
BBa_K3112041 | Composite | Chromosome stabilizing elements example 2 |
BBa_K3112042 | Composite | Chromosome stabilizing elements example 3 |
BBa_K3112043 | Composite | mRFP+HIS3 Chromosome Transfer Elements |
BBa_K3112044 | Composite | yEGFP+URA3 Chromosome Transfer Elements |
BBa_K3112045 | Composite | PGK1P+A to G adenine Base Editor+PGK1T |
BBa_K3112046 | Composite | Chromosomal stabilization element group example 3 |
Improved Parts
Name | Type | Description |
---|---|---|
BBa_K3112050 | Reporter | mRFP reporter with weak constitutive promoter(J23115) |
Necessary Parts
Bronze Standard
Name | Type | Description |
---|---|---|
BBa_E0422 | Reporter | ECFP (RBS+ LVA+ TERM) (B0034.E0022.B0015) |
BBa_E0430 | Reporter | EYFP (RBS+ LVA- TERM) (B0034.E0030.B0015) |
BBa_J23110 | Regulatory | constitutive promoter family member |
Silver Standard
Name | Type | Description |
---|---|---|
BBa_K3112041 | Composite | Chromosome stabilizing elements example 2 |
BBa_K3112042 | Composite | Chromosome stabilizing elements example 3 |
BBa_K3112043 | Composite | mRFP+HIS3 Chromosome Transfer Elements |
Gold Standard
Name | Type | Description |
---|---|---|
BBa_K3112050 | Reporter | mRFP reporter with weak constitutive promoter(J23115) |