Homology-directed genome editing is the intentional alteration of an endogenous genetic locus using information from an exogenous homology donor. A conversion tract is defined as the amount of genetic information that is converted from the homology donor to a given strand of the targeted chromosomal locus. Because of this, conversion tract analysis retrospectively not only elucidates the mechanism of homology-directed genome editing but also provides valuable insights on the conversion efficiency of every nucleotide in the homology donor. Here we describe a blue fluorescent protein-to-green fluorescent protein conversion system that can be conveniently used to measure the efficiency and analyze the lengths of conversion tracts of homology-directed genome editing using oligonucleotide donors in mammalian cells.
CITATION STYLE
Kan, Y., & Hendrickson, E. A. (2019). Conversion tract analysis of homology-directed genome editing using oligonucleotide donors. In Methods in Molecular Biology (Vol. 1999, pp. 131–144). Humana Press Inc. https://doi.org/10.1007/978-1-4939-9500-4_7
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