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Team:Paris Bettencourt/YonatanTest
From 2013.igem.org
Detect
Background
CRISPR/Cas systems generate site-specific double strand breaks and have recently been used for genome editing.
Aims
Building a genotype sensor based on CRISPR/Cas that reports existance of an antibiotic resistance gene.
Results
- Successfully cloned gRNA anti-KAN, crRNA anti-KAN, tracrRNA-Cas9 and pRecA-LacZ into Biobrick backbones and therefore generated four new BioBricks.
- Testing the new assembly standard for our cloning.
After expression of the Cas9 and gRNA, the gRNA guides the Cas9 to the target sequence, the kanamycin resistance. There, the Cas9 generates a double strand break. This activates the SOS response. The reporter LacZ is under the pRECA promoter, which gets activated during the SOS response and we get hence a blue cell, if the resistance gene has successfully been detected.
Centre for Research and Interdisciplinarity (CRI)
Faculty of Medicine Cochin Port-Royal, South wing, 2nd floor
Paris Descartes University
24, rue du Faubourg Saint Jacques
75014 Paris, France
+33 1 44 41 25 22/25
team2013@igem-paris.org
+33 1 44 41 25 22/25
team2013@igem-paris.org 
