Gene therapy progress and prospects: targeted gene repair

Parekh-Olmedo, Hetal; Ferrara, Luciana; Brachman, Erin E.; Kmiec, Eric B.

doi:10.1038/sj.gt.3302511

Cited by 65 publications

(69 citation statements)

References 50 publications

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“…17 According to the repair-dependent model ( Figure 1, Model I), pairing between the ssODN and its chromosomal complement leads to the formation of a displacement loop. DNA repair proteins would then introduce the desired base alteration into the chromosomal DNA strand using the ssODN as a template.…”

Section: Mechanistic Models For Ssodn-mediated Gene Targetingmentioning

confidence: 99%

Progress and prospects: oligonucleotide-directed gene modification in mouse embryonic stem cells: a route to therapeutic application

Aarts

Riele

2010

Gene Ther

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Gene targeting by single-stranded oligodeoxyribonucleotides (ssODNs) is a promising technique for introducing site-specific sequence alterations without affecting the genomic organization of the target locus. Here, we discuss the significant progress that has been made over the last 5 years in unraveling the mechanisms and reaction parameters underlying ssODN-mediated gene targeting. We will specifically focus on ssODN-mediated gene targeting in murine embryonic stem cells (ESCs) and the impact of the DNA mismatch repair (MMR) system on the targeting process. Implications of novel findings for routine application of ssODN-mediated gene targeting and challenges that need to be overcome for future therapeutic applications are highlighted.

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Section: Mechanistic Models For Ssodn-mediated Gene Targetingmentioning

confidence: 99%

Progress and prospects: oligonucleotide-directed gene modification in mouse embryonic stem cells: a route to therapeutic application

Aarts

Riele

2010

Gene Ther

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“…OMM makes use of different types of oligonucleotides: single-stranded DNA oligonucleotides containing 5 and/or 3 modified ends to protect the molecule against cellular nuclease activities (Campbell et al, 1989), chimeric RNA/DNA or DNA/DNA molecules D. Breyer et al (Igoucheva et al, 2004a;Parekh-Olmedo et al, 2005), RNA oligonucleotides (Storici, 2008), and triplexforming oligonucleotides (Simon et al, 2008).…”

Section: What Is Oligonucleotide-mediated Mutagenesis?mentioning

confidence: 99%

Commentary: Genetic modification through oligonucleotide-mediated mutagenesis. A GMO regulatory challenge?

Breyer

Herman

Brandenburger

et al. 2009

Environ. Biosafety Res.

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In the European Union, the definition of a GMO is technology-based. This means that a novel organism will be regulated under the GMO regulatory framework only if it has been developed with the use of defined techniques. This approach is now challenged with the emergence of new techniques. In this paper, we describe regulatory and safety issues associated with the use of oligonucleotide-mediated mutagenesis to develop novel organisms. We present scientific arguments for not having organisms developed through this technique fall within the scope of the EU regulation on GMOs. We conclude that any political decision on this issue should be taken on the basis of a broad reflection at EU level, while avoiding discrepancies at international level.

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“…The oligonucleotide must (1) enter the nucleus, (2) pair with its homologous target and (3) induce a base change at the targeted site via endogenous machinery. 12,13 It is not clear which of these barriers is most rate-limiting in ESC.…”

Section: Introductionmentioning

confidence: 99%

“…The ssODN may replace the target; it may induce a repair mechanism to alter the target strand; and/or it may become incorporated into one daughter cell during replication. 13 Gene correction may occur by more than one mechanism. There appears to be a 'strand bias' 12,16 in which targeting with antisense ssODNs often gives better results than sense ssODNs.…”

Section: Introductionmentioning

confidence: 99%

Delivery and mechanistic considerations for the production of knock-in mice by single-stranded oligonucleotide gene targeting

et al. 2006

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Single-stranded oligodeoxynucleotide (ssODN) gene targeting may facilitate animal model creation and gene repair therapy. Lipofection of ssODN can introduce point mutations into target genes. However, typical efficiencies in mouse embryonic stem cells (ESC) are o10 À4 , leaving corrections too rare to effectively identify. We developed ESC lines with an integrated mutant neomycin resistance gene (Tyr22Ter). After targeting with ssODN, repaired cells survive selection in G418. Correction efficiencies varied with different lipofection procedures, clonal lines, and ssODN designs, ranging from 1 to 100 corrections per million cells plated. Uptake studies using cell sorting of Cy5-labelled ssODN showed 40% of the corrections concentrated in the best transfected 22% of cells. Four different basepair mismatches were tested and results show that the basespecificity of the mismatch is critical. Dual mismatch ssODN also showed mismatch preferences. These ESC lines may facilitate development of improved ssODN targeting technologies for either animal production or ex vivo gene therapy.

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Gene therapy progress and prospects: targeted gene repair

Cited by 65 publications

References 50 publications

Progress and prospects: oligonucleotide-directed gene modification in mouse embryonic stem cells: a route to therapeutic application

Progress and prospects: oligonucleotide-directed gene modification in mouse embryonic stem cells: a route to therapeutic application

Commentary: Genetic modification through oligonucleotide-mediated mutagenesis. A GMO regulatory challenge?

Delivery and mechanistic considerations for the production of knock-in mice by single-stranded oligonucleotide gene targeting

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