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 - Jan 1 2019 |
Publication series
| Name | Methods in Molecular Biology |
|---|---|
| Volume | 1999 |
| ISSN (Print) | 1064-3745 |
| ISSN (Electronic) | 1940-6029 |
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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
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 proceeding › Chapter
}
TY - CHAP
T1 - Conversion tract analysis of homology-directed genome editing using oligonucleotide donors
AU - Kan, Yinan
AU - Hendrickson, Eric A
PY - 2019/1/1
Y1 - 2019/1/1
N2 - 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.
AB - 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.
KW - Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) (CRISPR-Cas9)
KW - Conversion tract
KW - Gene conversion
KW - Gene editing
KW - Homology-directed repair (HDR)
KW - Oligonucleotide
UR - http://www.scopus.com/inward/record.url?scp=85066290967&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85066290967&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-9500-4_7
DO - 10.1007/978-1-4939-9500-4_7
M3 - Chapter
T3 - Methods in Molecular Biology
SP - 131
EP - 144
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -