Contributions

The Future of Switch gRNA

We hope to provide a backbone for future iGEM research to expand upon the use of switch gRNA. The redesign of switch gRNA specific to c-Myc mRNA shows that switch gRNA can be realistically used and redesigned in novel research settings for iGEM teams. This opens the doors to iGEM teams re-designing switch gRNA to recognize different lengths of sequences, for example, allowing the detection of miRNA or larger sequences of mRNA. Conditional activation of CRISPR can also be used in tandem with another, different system that produces mRNA sequences to create a response. Switch gRNA is an incredible development from Collins et al. (2021) that we are grateful to have learned about and built upon by combining methods from Hmelo et al. (2015) and Taketo (1989). We look forward to introducing our work and its incredible potential to the iGEM community.

References

Collins, S. P.; Rostain, W.; Liao, C.; Beisel, C. L. Sequence-Independent RNA Sensing and DNA Targeting by a Split Domain CRISPR–Cas12a gRNA Switch. Nucleic acids research 2021, 49 (5), 2985–2999. https://doi.org/10.1093/nar/gkab100.

Hmelo L. R.; Borlee B. R.; Almblad H.; Love M. E.; Randall T. E.; Tseng B. S.; Lin C.; Irie Y.; Storek K. M.; Yang J. J.; Siehne R. J.; Howell P. L.; Singh P. K.; Tolker-Nielsen T.; Parsek M. R.; Schweizer H. P.; Harrison J. J. Precision-Engineering the Pseudomonas Aeruginosa Genome with Two-Step Allelic Exchange. Nature Protocols [Online] 2015, 10:1820–1841. https://doi.org/10.1038/nprot.2015.115.

Taketo, A. RNA Transfection of Escherichia Coli by Electroporation. Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression [Online] 1989, 1007:127–129. https://doi.org/10.1016/0167-4781(89)90142-5.