DNA oligonucleotides and plasmids perform equally as donors for targeted gene conversion.

Abstract:

:Site-specific gene modifications in cells are initiated by the introduction of exogenous DNA. We used a recently established cell assay to compare the ability of DNA donors to induce a single point mutation that converts a target gene encoding blue fluorescent protein (BFP) into expressing green fluorescent protein (GFP). In a chromosomal assay with cells stably expressing BFP, we showed that fluorescently labeled single-stranded oligonucleotides and a donor plasmid cotranscribing a red fluorescent protein provide similar efficiencies in triggering BFP-GFP conversions. In transient cotransfections, an isogenic donor plasmid comprising a nonfunctional GFP gene yielded a greater efficiency for the conversion of the BFP target gene than a nonisogenic donor, and all plasmid donors were superior to oligonucleotides.

journal_name

Biochem Genet

journal_title

Biochemical genetics

authors

Wagner S,McCracken J,Cole S,Laible G

doi

10.1007/s10528-010-9370-z

subject

Has Abstract

pub_date

2010-12-01 00:00:00

pages

897-908

issue

11-12

eissn

0006-2928

issn

1573-4927

journal_volume

48

pub_type

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