Conversion tract analysis of homology-directed genome editing using oligonucleotide donors

Research output: Chapter in Book/Report/Conference proceedingChapter

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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 languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages131-144
Number of pages14
DOIs
StatePublished - 2019

Publication series

NameMethods in Molecular Biology
Volume1999
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

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

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