Team:Tec-Monterrey/Description

CRISPR-Cas technology has the capability of storing information. This year, iGEM team Tec-Monterrey aims to use the CRISPR-Cas system to store specific DNA sequences in the genome of E. coli in order to save information about the environment surrounding the bacteria. To make this possible, Cas1-Cas2 proteins, which create the protospacer acquisition in the CRISPR system, are used to insert a synthetic DNA sequence in the CRISPR array within the genome of the bacteria. This synthetic sequence is produced by a second system, called SCRIBE. The final step of our project is reading out the inserted DNA sequence. Using specific primers for polymerase chain reaction (PCR) are used to amplify a section of the CRISPR array where the sequence is inserted. Taking together both systems, our project intends to act as a biological tape recorder capable of sensing external stimuli in the environment and storing their presence in the genome.

References
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Díez-Villaseñor, C., Guzmán, N. M., Almendros, C., García-Martínez, J. & Mojica, F. J. M. CRISPR-spacer integration reporter plasmids reveal distinct genuine acquisition specificities among CRISPR-Cas I-E variants of Escherichia coli. RNA Biol. (2013). doi:10.4161/rna.24023

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Nuñez, J. K., Kranzusch P, Noeske J, Wright A, Davies C, Doudna J. Cas1-Cas2 complex formation mediates spacer acquisition during CRISPR-Cas adaptive immunity. Nat. Struct. Mol. Biol. (2014). doi:10.1038/nsmb.2820

Sheth, R. U., Yim, S. S., Wu, F. L. & Wang, H. H. Multiplex recording of cellular events over time on CRISPR biological tape. Science. (2017). doi:10.1126/science.aao0958

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Tang, W., & Liu, D. R. Rewritable multi-event analog recording in bacterial and mammalian cells. Science. (2018). doi: 10.1126/science.aap8992

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