Protein Sequences. The Genetic Code The natural extension of the genetic code…
Rewriting the Genetic Code
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Transcript of Rewriting the Genetic Code
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Rewriting the Genetic Code
By:Ishika Desai
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Genetic Code
• DNA is made up of long sequences of letters that code for specific amino acids.
• Codons are made up of three of these nucleotides (ACTG)
• Most three letter combos code for amino acids, but some represent stop codons.
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How does it work?
• System overwrites specific DNA sequences within the genome
• Uses Multiplex Automated Genome Engineering (MAGE) to site-specifically replace stop codons across E. Coli strains
• Uses Conjugative Assembly Genome Engineering (CAGE) to precisely control a naturally occurring process that bacteria use to exchange genetic material
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Potential Advances
• Engineer bacteria that are resistant to several viruses
• Build proteins not found in nature
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Bibliography• Isaacs, Farren J., Harris H. Wang, and Peter A. Carr. "Precise
Manipulation of Chromosomes in Vivo Enables Genome-Wide Codon Replacement." Science Magazine: Sign In. N.p., 15 July 2011. Web. 09 July 2013. <http://www.sciencemag.org/content/333/6040/348.full>.
• Trafton, Anne. "Scientists Unveil Tools for Rewriting the Codeof Life." MIT's News Office. N.p., 15 July 2011. Web. 09 July 2013. <http://web.mit.edu/newsoffice/2011/editing-genome-0715.html>.
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MedulloBLASToma
By: Ishika Desai
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Medulloblastoma
• Malignant brain tumor located in the cerebellum
• Makes up 20% of brain tumors in children• Cause of medulloblastoma is unknown• Can become cancerous if it is combined with
the expression of the catenin beta 1 gene
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β-catenin Gene
• Officially known as CTNNB1 • Expression of β-catenin is associated with:
• Carcinoma• Lung Cancer• Medulloblastoma• Malignant breast tumors• Endometrial cancer• Ovarian cancer
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shRNA is generated within the aerotolerant E. coli via RNA interference
E. coli binds to β-1 integrin receptors on the cancerous cells and invade them
shRNA segments bind to β-catenin mRNA transcripts to silence the expression of this gene
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shRNA production within E. coli
Pol II
Pre-
miR
NA
shRNA-induced silencing complex
Drosha
shRN
A
lysteriolysin O
Exportin 5 Dicer enzyme
Leaves E. coli
DNA
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shRNA
shRNA
≈≈≈INVASIN PRODUCTION
GENETICALLY ENGINEERED E. COLI
CANCEROUS CELL
CTNNB1 mRNA
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• Methods currently used to treat cancerous medulloblastomas:
• Surgery• Radiation • Chemotherapy
• All of the methods listed above generally still result in a considerable amount of damage to healthy tissue
• Oncologists and researchers have been looking into cancer treatments that precisely distinguish between healthy and diseased cells
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Expected ResultsshRNA from E. coli Expression of CTNNB1
0 1
1 0
A XInverter (NOT) Logic Gate
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Expected Results
• After invading the cancerous cells, it is expected that the shRNA in the RISC will bind to the CTNNB1 mRNA transcripts and induce mRNA cleavage
• This will silence the CTNNB1 gene