Team:Danci-K8/Results

Overview

We generate a SNPs gene panel which can determine people's preference of food tastes using Taqman assay on the Fluidigm nano-fluidic gene dynamic array chips (96:96). Endpoint fluorescent image data is acquired on the BioMark™ System for genetic analysis and data is analyzed using the Fluidigm SNP Genotyping Analysis software, to obtain genotype calls. In addition to saliva collection from students volunteering to participate, after completing their phenotypes forms of response to food tastes, we generate a synthetic DNA harboring the wildtype (taste) and mutant alleles (non-taste) (SNP) for the ability to sense taste which will be used as control for the Taqman assays and for preforming CRISPR on the mutant SNP allele, to convert it to the wildtype allele, as a feasibility test to be able to change people's taste sense and help them to consume necessary oral medications even if they taste very bitter to them.

Registry parts (Biobricks)

Fluidigm gene chip array

Key Achievements:
  1. We created a genetic toolbox consisting of 5 knowing taste with 30 SNPs Including the wild type (taste) and mutant (non-taste). This the first genomes sequenced in the history of iGEM.
  2. We generate a SNPs gene panel which can determine people's preference of food tastes. This the first panel which includes a broad library that contains all five tastes of human.

SNPs gene panel-which can determine people's preference of food tastes using Taqman assay

NaCl & Sour taste

Fig 1- Results of DNA students samples NaCl & Sour taste, Compared to synthetic SNP DNA in the panel in Taqman reaction conditions on the Fluidigm chip. Wild Type (Taste SNPs) marked with VIC- green color. Mutant (Non-taste SNPs) is marked with FAM- red color. Blue color (show with green and red color) - mean that there are population genetic polymorphisms in the country.

Fig 2- The population genetic polymorphism in the country in the context of this SNP.

Umami & Monosodium gluten

Fig 3- Results of DNA students samples Umami & Monosodium gluten taste Compared to synthetic SNP DNA in the panel in Taqman reaction conditions on the Fluidigm chip. Mutant (Non-taste SNPs) is marked with FAM- red color. We can see that there are no population genetic polymorphisms in the country of this SNP.

Fig 4- The population genetic polymorphism in the country in the context of this SNP.

Cilantro preference

Fig 5- Results of DNA students samples Cilantro preference Compared to synthetic SNP DNA in the panel in Taqman reaction conditions on the Fluidigm chip. Wild Type (Taste SNPs) marked with VIC- green color. Mutant (Non-taste SNPs) is marked with FAM- red color. Blue color (show with green and red color) - mean that there are population genetic polymorphisms in the country.

Fig 6- The population genetic polymorphism in the country in the context of this SNP.

Sweet

Fig 7- Results of DNA students samples Sweet taste Compared to synthetic SNP DNA in the panel in Taqman reaction conditions on the Fluidigm chip. Wild Type (Taste SNPs) marked with VIC- green color. Mutant (Non-taste SNPs) is marked with FAM- red color. Blue color (show with green and red color) - mean that there are population genetic polymorphisms in the country.

Fig 8- The population genetic polymorphism in the country in the context of this SNP.

Grapefruit juice bitterness

Fig 9- Results of DNA students samples Grapefruit juice bitterness taste Compared to synthetic SNP DNA in the panel in Taqman reaction conditions on the Fluidigm chip. Wild Type (Taste SNPs) marked with VIC- green color. Mutant (Non-taste SNPs) is marked with FAM- red color. Blue color (show with green and red color) - mean that there are population genetic polymorphisms in the country.

Fig 10- The population genetic polymorphism in the country in the context of this SNP.

Consuming Alcoholic

Fig 11- Results of DNA students samples Consuming Alcoholic Compared to synthetic SNP DNA in the panel in Taqman reaction conditions on the Fluidigm chip. Wild Type (Taste SNPs) marked with VIC- green color. Mutant (Non-taste SNPs) is marked with FAM- red color. Blue color (show with green and red color) - mean that there are population genetic polymorphisms in the country.

Fig 12- The population genetic polymorphism in the country in the context of this SNP.

PTC

Fig 13- Results of DNA students samples PTC Compared to synthetic SNP DNA in the panel in Taqman reaction conditions on the Fluidigm chip. Wild Type (Taste SNPs) marked with VIC- green color. Mutant (Non-taste SNPs) is marked with FAM- red color. Blue color (show with green and red color) - mean that there are population genetic polymorphisms in the country.

Fig 14- The population genetic polymorphism in the country in the context of this SNP.

Coffee bitterness

Fig 15- Results of DNA students samples Coffee bitterness Compared to synthetic SNP DNA in the panel in Taqman reaction conditions on the Fluidigm chip. Wild Type (Taste SNPs) marked with VIC- green color. Mutant (Non-taste SNPs) is marked with FAM- red color. Blue color (show with green and red color) - mean that there are population genetic polymorphisms in the country.

Fig 16- The population genetic polymorphism in the country in the context of this SNP.

CRISPR cas9

Key Achievements
  1. We generate 4 specific guide RNA to be able to change people's taste sense.
  2. Guide RNA consist to : Coffee bitterness, Grapefruit juice bitterness, Umami & Monosodium gluten, NaCl & Sour tastes.

Fig 17
1-4 - Mutant alleles (non-taste) Cutting with CRISPR cas9.
5- Non target gRNA - Negative control.
6- PUC57 plasmid

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