problem
With more than
77,000 new cases every year,
B-cell lymphoma is a type of Non-Hodgkin's lymphoma that reduces white blood cells' ability to fight infections.
This blood cancer specifically kills more than
1 in every 100,000 people.
It originates in lymphocytes found in the lymph nodes, spleen, and other secondary lymphoid organs.
problem
current diagnosis
Current diagnosis procedures for B-cell lymphoma include biopsies, gene expression profiling, and PET imaging Immunophenotyping and flow cytometry can cost up to $100,000. The cost and invasiveness of these methods mean they aren't utilized until symptoms have developed. Thus, early and inexpensive diagnostics are crucial to manage this cancer.
We aim to equip E.coli with a genetically engineered gene circuit to develop a less expensive detection method for clinical use.
B-cell lymphoma will be detected by the elevated presence of microRNA, specifically miRNA-326.With our engineered E.coli in a microfluidic assay, only a small blood sample will be needed.
Solution
Our bacterial biosensor will detect microRNA with the integration of 2 plasmids: one of human Argonaute 2 (Ago2),and the other of a dual repression system: transcriptional repressor LacI, translational repressor L7Ae, and GFP
Without miRNA, LacI and L7Ae repress GFP. However, with miRNA, a RNA-induced silencing complex (RISC) is formed with Ago2. The RISC cleaves the target site on the repressors' mRNA that is complementary to the miRNA guide strand. Thus, LacI and L7Ae no longer repress GFP, and the biosensor can fluoresce.
our
engineering
broader
application
We aim to transform miRNA into the competent cells of our engineered E.coli, a relatively novel and undocumented process. Success with this would be a significant contribution to research surrounding miRNA and RNA activity in bacteria.
This circuit can be adapted to detect virtually any miRNA by switching out different target sequences. Additionally, this detection can be refined by adding in more miRNA target sequences that are specific to the disease.
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Project Description
Wet Lab Result
Model
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