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

Research output: Chapter in Book/Report/Conference proceedingChapter

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 - Jan 1 2019

Publication series

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

Fingerprint

Oligonucleotides
Genetic Loci
Green Fluorescent Proteins
Nucleotides
Gene Editing

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

PubMed: MeSH publication types

  • Journal Article

Cite this

Kan, Y., & Hendrickson, E. A. (2019). Conversion tract analysis of homology-directed genome editing using oligonucleotide donors. In Methods in Molecular Biology (pp. 131-144). (Methods in Molecular Biology; Vol. 1999). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-9500-4_7

Conversion tract analysis of homology-directed genome editing using oligonucleotide donors. / Kan, Yinan; Hendrickson, Eric A.

Methods in Molecular Biology. Humana Press Inc., 2019. p. 131-144 (Methods in Molecular Biology; Vol. 1999).

Research output: Chapter in Book/Report/Conference proceedingChapter

Kan, Y & Hendrickson, EA 2019, Conversion tract analysis of homology-directed genome editing using oligonucleotide donors. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 1999, Humana Press Inc., pp. 131-144. https://doi.org/10.1007/978-1-4939-9500-4_7
Kan Y, Hendrickson EA. Conversion tract analysis of homology-directed genome editing using oligonucleotide donors. In Methods in Molecular Biology. Humana Press Inc. 2019. p. 131-144. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-9500-4_7
Kan, Yinan ; Hendrickson, Eric A. / Conversion tract analysis of homology-directed genome editing using oligonucleotide donors. Methods in Molecular Biology. Humana Press Inc., 2019. pp. 131-144 (Methods in Molecular Biology).
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