High-frequency genome editing using ssDNA oligonucleotides with zinc-finger nucleases

Fuqiang Chen; Shondra M Pruett-Miller; Yuping Huang; Monika Gjoka; Katarzyna Duda; Jack Taunton; Trevor N Collingwood; Morten Frödin; Gregory D Davis

doi:10.1038/nmeth.1653

High-frequency genome editing using ssDNA oligonucleotides with zinc-finger nucleases

Fuqiang Chen, Shondra M Pruett-Miller, Yuping Huang, Monika Gjoka, Katarzyna Duda, Jack Taunton, Trevor N Collingwood, Morten Frödin, Gregory D Davis

316 Citations (Scopus)

Abstract

Zinc-finger nucleases (ZFNs) have enabled highly efficient gene targeting in multiple cell types and organisms. Here we describe methods for using simple ssDNA oligonucleotides in tandem with ZFNs to efficiently produce human cell lines with three distinct genetic outcomes: (i) targeted point mutation, (ii) targeted genomic deletion of up to 100 kb and (iii) targeted insertion of small genetic elements concomitant with large genomic deletions.

Original language	English
Journal	Nature Methods
Volume	8
Issue number	9
Pages (from-to)	753-5
Number of pages	3
DOIs	https://doi.org/10.1038/nmeth.1653
Publication status	Published - Sept 2011

Keywords

Cell Line
DNA, Single-Stranded
Endonucleases
Gene Deletion
Gene Targeting
Genetic Engineering
Humans
Mutagenesis, Insertional
Oligodeoxyribonucleotides
Point Mutation
Zinc Fingers

Access to Document

10.1038/nmeth.1653

Cite this

@article{a8272182525f4499b6ae79ea7adfbb8f,

title = "High-frequency genome editing using ssDNA oligonucleotides with zinc-finger nucleases",

abstract = "Zinc-finger nucleases (ZFNs) have enabled highly efficient gene targeting in multiple cell types and organisms. Here we describe methods for using simple ssDNA oligonucleotides in tandem with ZFNs to efficiently produce human cell lines with three distinct genetic outcomes: (i) targeted point mutation, (ii) targeted genomic deletion of up to 100 kb and (iii) targeted insertion of small genetic elements concomitant with large genomic deletions.",

keywords = "Cell Line, DNA, Single-Stranded, Endonucleases, Gene Deletion, Gene Targeting, Genetic Engineering, Humans, Mutagenesis, Insertional, Oligodeoxyribonucleotides, Point Mutation, Zinc Fingers",

author = "Fuqiang Chen and Pruett-Miller, {Shondra M} and Yuping Huang and Monika Gjoka and Katarzyna Duda and Jack Taunton and Collingwood, {Trevor N} and Morten Fr{\"o}din and Davis, {Gregory D}",

year = "2011",

month = sep,

doi = "10.1038/nmeth.1653",

language = "English",

volume = "8",

pages = "753--5",

journal = "Nature Methods",

issn = "1548-7105",

publisher = "nature publishing group",

number = "9",

}

TY - JOUR

T1 - High-frequency genome editing using ssDNA oligonucleotides with zinc-finger nucleases

AU - Chen, Fuqiang

AU - Pruett-Miller, Shondra M

AU - Huang, Yuping

AU - Gjoka, Monika

AU - Duda, Katarzyna

AU - Taunton, Jack

AU - Collingwood, Trevor N

AU - Frödin, Morten

AU - Davis, Gregory D

PY - 2011/9

Y1 - 2011/9

N2 - Zinc-finger nucleases (ZFNs) have enabled highly efficient gene targeting in multiple cell types and organisms. Here we describe methods for using simple ssDNA oligonucleotides in tandem with ZFNs to efficiently produce human cell lines with three distinct genetic outcomes: (i) targeted point mutation, (ii) targeted genomic deletion of up to 100 kb and (iii) targeted insertion of small genetic elements concomitant with large genomic deletions.

AB - Zinc-finger nucleases (ZFNs) have enabled highly efficient gene targeting in multiple cell types and organisms. Here we describe methods for using simple ssDNA oligonucleotides in tandem with ZFNs to efficiently produce human cell lines with three distinct genetic outcomes: (i) targeted point mutation, (ii) targeted genomic deletion of up to 100 kb and (iii) targeted insertion of small genetic elements concomitant with large genomic deletions.

KW - Cell Line

KW - DNA, Single-Stranded

KW - Endonucleases

KW - Gene Deletion

KW - Gene Targeting

KW - Genetic Engineering

KW - Humans

KW - Mutagenesis, Insertional

KW - Oligodeoxyribonucleotides

KW - Point Mutation

KW - Zinc Fingers

U2 - 10.1038/nmeth.1653

DO - 10.1038/nmeth.1653

M3 - Journal article

C2 - 21765410

SN - 1548-7105

VL - 8

SP - 753

EP - 755

JO - Nature Methods

JF - Nature Methods

IS - 9

ER -

High-frequency genome editing using ssDNA oligonucleotides with zinc-finger nucleases

Abstract

Keywords

Access to Document

Fingerprint

Cite this