Bloom Busters Project Contributions

In our project, our team had the goal of genetically engineering Chlamydomonas reinhardtii to increase its ability to sequester phosphorous, and to increase its uptake of nitrate by converting nitrous oxide to dinitrogen gas.

To accomplish this, our team developed 3 new simple parts. We utilized additional simple parts from the Chlamdydomonas reinhardtii MoClo toolkit (Crozet et al. 2016), which have been documented by other iGEM Teams, along with a simple part for the Psr1 gene from the Alison Baker Lab (Slocombe et al. 2023). This led to the generation of 8 new composite parts. A brief description of these parts is in the table below.

For more information, including a description of which simple parts were used to assemble each composite part, please visit the part’s Registry page!

part Chlamy MoChlo Toolkit # Registry Number iGEM Team/Collaborator Part Description/Purpose
nanoLuc PCM0-104 K2619004 iGEM18_XJTLU-CHINA luciferase reporter
aadA CDS PCM0-076 K3002000 iGEM19_TU_Kaiserslautern spectinomycin resistance
Psad Promoter+5' UTR PCM0-016 K3002001 iGEM19_TU_Kaiserslautern Strong promoter in C. reinhardtii
Psad Terminator PCM0-114 K3002002 iGEM19_TU_Kaiserslautern terminator in C. reinhardtii
Btub promoter+5' UTR PCM0-021 K3002011 iGEM19_TU_Kaiserslautern Strong promoter in C. reinhardtii
Btub Terminator PCM0-118 K3002012 iGEM19_TU_Kaiserslautern terminator in C. reinhardtii
mCherry PCM0-108 K4770013 iGEM23_Barcelona-UB fluorescent reporter
NosZ P. stutzeri K5078000 new nitrous oxide reductase from Pseudomonas stutzerii
NosZ D. denitirficans K5078001 new nitrous oxide reductase from Dechloromonas denitrificans
Ptc amiRNA K5078002 new encodes miRNA against Ptc mRNA
Psr gDNA K5078008 provided by Alison Baker Lab (Slocombe et al. 2023) Phosphate starvation response 1 transcription factor
pL1-aadA K5078004 new Composite part - provide C. reinhardtii with resistance to spectinomycin
pL1 NosZ (Pstu)-mCherry K5078005 new Composite part - allow C. reinhardtii to convert nitrous oxide to dinitrogen gas, tagged with mCherry reporter
pL1 NosZ (D denit)-mCherry K5078006 new Composite part - allow C. reinhardtii to convert nitrous oxide to dinitrogen gas, tagged with mCherry reporter
pL1 ptc amiRNA K5078007 new Composite part - reduce expression of Ptc mRNA
pL1 Psr1-nanoLuc K5078003 new Composite part - upregulate phosphate starvation response in C. reinhardtii, tagged with nanoLuc reporter
pL2 -aadA-nosZ(P.stu)-Ptc amiRNA-Psr K5078009 new Composite part - combination of above pL1 functions
pL2 -aadA-nosZ(D.denit)-Ptc amiRNA-Psr K5078010 new Composite part - combination of above pL1 functions
pL2-aadA-Psr K5078011 new Composite part - combination of above pL1 functions