Team:Shenzhen BGIC ATCG/stories

The Magic

This year, our project named "Cell Magic". We engineered the budding yeast in time level as well as space level. "Cell Magic", like a movie,can be divided into two parts: the actor related and the time regulation related one. In the first part, our actor, signal peptides, were all made up with colorful clothes by fluorescent proteins. Furthermore, the intron and degradation biobricks as make-up artists can decide how the clothes be matched. Through these steps, our actors may wearing in green, yellow or even mixed, appear in their specific sub-locations of yeast cell. With respect to the time level, we took advantages of the natural cell cycle in budding yeast to improve the time process to be more suitable for our movie.In general, the movie director, the promoters from five cycline gene, can decide the expression of downstream gene in G1 and G2 phase phase, respectively. Also, freezer Sic1 help our movie stay a longer time in the G1 phase which is important for the actor performing their stories. And the last tools we utilized is the microfluidics, through which numerous cells can project our "Cell Magic", thus ensuring our observation by naked eyes.

Cyclin Promoters

Cell cycle is a complex process and can be separated into G0, G1, S, G2, M phase. In each phase, distinct transcription factors help the phase-specific gene express through recognizing their promoters. Thus, these promoters are phase-specific too and can be fused with other genes in order to express such gene in a defined cell cycle phase. As we all know, different proteins are planned to be expressed along the cell cycle. And their mRNAs are transcripted through the specific promoters. These promoters locate at their upstream sequences and can be recognized by the RNA polymerase, which can initiate such process. There are databases containing the upstream 600 amino acid sequence in the upstream of DNA, which can be transcript in the G1, S, G2 and M phase, respectively [http://www.genome.jp/kegg-bin/show_pathway?org_name=sce&mapno=04111&mapscale=&show_description=hide]. Previous study 【Candidate regulatory sequence elements for cell】demonstrated that cell cycle specific promoters posses their conserved five to six base pairs. Thus, we found the high-confidence 600 promoter-containing sequences in database that harbor the paper-mentioned 5/6 bp sequence. Consequently, we pick up the upstream 600bp sequence of cln2, cln3, clb2, clb5 and clb6. The Clb2 gene is highly expressed in G2 phase, and the genes are very strongly induced by GAL-CLB2, whereas GAL-CLN3 appears somewhat repressive 【Comprehensive Identification of Cell Cycle–regulated Genes of the Yeast Saccharomyces cerevisiae by Microarray Hybridization】

Reporter Modification

Degradation Tags

Targeting Peptides

Cell Cycle Regulator

Alternative Splicing Device

Microfluidic Device

Cell Synchronization