Team:OUC-China/Proof
NATIVE STEM-LOOPS
I. Validate that native stem-loops can affect the expression of its flanking cistron based on our platform Click here to know more.
Figure.1 Both relative expression on RNA and protein level with stem-loop of -25.6 kcal/mol (measured by Mfold) are significantly higher than those of control group without stem-loop, which means stem-loops can coordinate the expression within cistrons. The vertical coordinate is the ratio of upstream gene gfp to downstream gene mcherry. Error bars indicate s.d. of mean of experiments in triplicate.
II. Prove that native stem-loops with gradient folding free energy results in differential expression. Click here to know more.
Figure.2 Stem-loops with lower folding free energy lead to stronger protection of upstream gene, which results in higher ratio of GFP to mCherry both in transcriptional and translational level. Error bars indicate s.d. of mean of experiments in triplicate.
DESIGNED STEM-LOOPS
III. Verify the regulatory effect of designed stem-loops.
Figure.3 One of the designed stem-loop of -34.4 kcal/mol (measured by Mfold) inserted into intergenic regions can also tune the expression of gfp and mcherry as expected, similarly with the effect of native stem-loops. Error bars indicate s.d. of mean of experiments in triplicate.
STANDARDIZE PARTS
IV. Standardize measured regulatory elements and submit them to iGEM parts registry with detailed characterization. See more at parts.
CONSTRUCT TOOLKIT
V. Constructed a set of toolkit of stem-loops with different folding free energy.
Figure.4 Detailed data of designed stem-loops with various folding free energy in our toolkit. Click here to know more.
MODEL
VI. Model the process of mRNA degradation in the present of stem-loops. See more at model.
SOFTWARE
VII. Create software to realize sequence scanning and stem-loop predicting based on our model. See more at software.
Cistrons Concerto
Thanks
1.Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences
2.NEW ENGLAND Biolabs
3.GenScript
Contact us:
E-mail: oucigem@163.com
Designed and built by @ Jasmine Chen and @ Zexin Jiao
We are OUC-iGEM
