Abstract
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.
| Original language | English (US) |
|---|---|
| Title of host publication | Methods in Molecular Biology |
| Publisher | Humana Press Inc. |
| Pages | 131-144 |
| Number of pages | 14 |
| DOIs | |
| State | Published - 2019 |
Publication series
| Name | Methods in molecular biology (Clifton, N.J.) |
|---|---|
| Publisher | Humana Press |
| ISSN (Print) | 1064-3745 |
Bibliographical note
Publisher Copyright:© Springer Science+Business Media, LLC, part of Springer Nature 2019.
Keywords
- Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) (CRISPR-Cas9)
- Conversion tract
- Gene conversion
- Gene editing
- Homology-directed repair (HDR)
- Oligonucleotide