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

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10.1038/nmeth.1653

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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",

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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

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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

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