Gene editing depends upon the cell’s endogenous DNA repair pathways. Upon DNA double-strand break, two known repair pathways are enacted: nonhomologous end-joining (NHEJ) and homologous recombination (HDR), among others. The HDR pathway works only in the presence of an exogenous homologous template. Is it possible to determine which pathway is activated in a cell after gene disruption (GD)? Is it NHEJ or HDR? Certo., et al. 2011 [1] published an article on the Traffic Light Reporter (TLR) that can distinguish between NHEJ and HR events. Traffic light reporter system is designed such that cells generate flow-cytometric readout of HDR-mediated gene targeting or mutagenic NHEJ-mediated gene disruption occurring at an individual DNA breakpoint, which is extremely exciting!

References

1. Michael T Certo., et al. “Tracking genome engineering outcome at individual DNA breakpoints”. Nature Methods 8 (2011): 671-676.
2. Ryan Kuhar., et al. “Novel fluorescent genome editing reporters for monitoring DNA repair pathway utilization at endonuclease-induced breaks”. Nucleic Acids Research 42.1 (2014): e4.

Citation

Citation: Kawalpreet K Aneja. “Traffic Light Reporter for Genome Engineering". Acta Scientific Microbiology 3.9 (2020): 27-28.

Copyright

Copyright: © 2020 Kawalpreet K Aneja. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.