Part:BBa_K5242015
ogRNA_GLC3
Biology of GLC3
GLC3 is the gene encoding 1,4-α-glucan-branching enzyme in Saccharomyces cerevisiae. The gene is located at V:133,120 - 135,234 and is involved in the glycogen biosynthesis process. [1]It adds branches to glycogen molecules. [2]GLC3 mRNA begins to accumulate when environmental glucose decreases to approximately 50% and peaks when environmental glucose is depleted, similar to other glycogen metabolism genes.[3]
Usage
This part can be complementarily paired with GLC3 and contains an A-C mismatch site in the middle for the introduction of ADAR editing. And this part is the core part of the sensor RNA used to detect Chk1. The addition of four of these MS2 sequences facilitates ADAR recognition and editing. At the beginning and end of the fragment, we added E2A and LV2A peptides, respectively, allowing both fluorescent proteins to be separated from the intervening peptide throughout sensor RNA translation.
Figure.1 shows the structure of our Strain Security System. If we choose GLC3 as the monitoring object, then the ogRNA in the figure refers to ogRNA_GLC3. Under specific conditions like heat stress, the concentration of stress response gene transcripts is high, which can form duplex with sensor. Thus, ADAR is able to edits the stop codon of the ogRNA sequence in the sensor, allowing for the expression of Cre recombinase. The Cre recombinase can knockout the suicide gene, preventing the death of the strain.
When conditions change, Cre recombinase is not expressed in large quantities, so the suicide gene is continuously expressed and accumulates, eventually leading to the death of the strain. This method avoids the drawbacks of persistent leakage expression of the suicide gene, which could otherwise damage the cells.Figure.1 shows the structure of our Strain Security System.
Figure 1.The design of Strain Security System
Referances
[1]UniProt: the Universal Protein Knowledgebase in 2023
[2]Thon, V. J.et al. Coordinate regulation of glycogen metabolism in the yeast Saccharomyces cerevisiae. Induction of glycogen branching enzyme.Journal of biological Chemistry 267, 15224-15228 (1992).
[3] Parrou, J. L.et al. Dynamic responses of reserve carbohydrate metabolism under carbon and nitrogen limitations in Saccharomyces cerevisiae.Yeast 15, 191-203 (1999). Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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