High-level expression of hemoglobin A in human thalassemic erythroid progenitor cells following lentiviral vector delivery of an antisense snRNA

Vacek, Marla M.; Ma, Hong; Gemignani, Federica; Lacerra, Giuseppina; Kafri, Tal; Kole, Ryszard

doi:10.1182/blood-2002-06-1869

Cited by 45 publications

(32 citation statements)

References 52 publications

(64 reference statements)

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“…4A and inset). As was the case for AONs, such modified U7 snRNAs were initially almost exclusively designed to induce exon skipping in genes involved in human diseases such as β-thalassemia, [134][135][136][137] Duchenne muscular dystrophy, [138][139][140] or HIV/AIDS. [141][142][143] Focusing on SMA, a number of different U snRNA-based approaches have been used with variable success (Fig.…”

Section: Smn2 Exon 7 Inclusionmentioning

confidence: 99%

Repair of pre-mRNA splicing

2010

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Section: Smn2 Exon 7 Inclusionmentioning

confidence: 99%

Repair of pre-mRNA splicing

2010

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“…Another β-globin mutation, β E , was also successfully targeted using this approach (6). In an alternative and potentially longer-term strategy, the antisense sequences were incorporated into lentiviral expression vectors and used to successfully treat human erythropoietic progenitor cells ex vivo (7).…”

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confidence: 99%

Therapeutic potential of antisense oligonucleotides as modulators of alternative splicing

Sazani¹,

Kole²

2003

J. Clin. Invest.

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111

102

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Correction of splicing by antisense oligonucleotides β-Globin. β-Thalassemia is a serious genetic blood disorder (3) in which production of β-globin, a subunit of hemoglobin, is partially or entirely ablated by mutations in the β-globin gene. The defect decreases the oxygen-carrying capacity of red blood cells and causes compensatory expansion of the bone marrow An estimated 60% of all human genes undergo alternative splicing, a highly regulated process that produces splice variants with different functions. Such variants have been linked to a variety of cancers, and genetic diseases such as thalassemia and cystic fibrosis. This Perspective describes a promising approach to RNA repair based on the use of antisense oligonucleotides to modulate alternative splicing and engender the production of therapeutic gene products.

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“…From our experiment, intronic cryptic donor sites are relatively easy to suppress by U7 snRNAs. In contrast, although more cases need to be investigated, the suppression of exonic cryptic sites may result in skipping of the exon in which they are located depending on the presence of the nearby ESE as in CYP11A 566C [ T. Exon skipping in a similar situation has been reported by Vacek et al (2003). They have used a U7 snRNA targeting an exon-internal location to induce exon skipping in a thalassemic HBB gene.…”

Section: Discussionmentioning

confidence: 92%

“…Of note, oligonucleotide with a two-nucleotide mismatch gave no antisense effect (Sierakowska et al 1996). Second, neither of the plasmids caused any detectable changes in the morphology or growth rate of transfected cells (Sierakowska et al 1996;Vacek et al 2003; data not shown). However, it may be intriguing to see the influence of engineered U7 snRNA on cellular splicing using exon junction microarrays (Johnson et al 2003;Nagao et al 2005a).…”

Section: Discussionmentioning

confidence: 99%

U7 snRNA-mediated correction of aberrant splicing caused by activation of cryptic splice sites

et al. 2007

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A considerable fraction of mutations associated with hereditary disorders and cancers affect splicing. Some of them cause exon skipping or the inclusion of an additional exon, whereas others lead to the inclusion of intronic sequences or deletion of exonic sequences through the activation of cryptic splice sites. We focused on the latter cases and have designed a series of vectors that express modified U7 small nuclear RNAs (snRNAs) containing a sequence antisense to the cryptic splice site. Three cases of such mutation were investigated in this study. In two of them, which occurred in the PTCH1 and BRCA1 genes, canonical splice donor sites had been partially impaired by mutations that activated nearby intronic cryptic splice donor sites. Another mutation found in exonic region in CYP11A created a novel splice donor site. Transient expression of the engineered U7 snRNAs in HeLa cells restored correct splicing in a sequence-specific and dosedependent manner in the former two cases. In contrast, the third case, in which the cryptic splice donor site in the exonic sequence was activated, the expression of modified U7 snRNA resulted in exon skipping. The correction of aberrant splicing by suppressing intronic cryptic splice sites with modified U7 is expected be a promising alternative to gene replacement therapy.

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High-level expression of hemoglobin A in human thalassemic erythroid progenitor cells following lentiviral vector delivery of an antisense snRNA

Cited by 45 publications

References 52 publications

Repair of pre-mRNA splicing

Repair of pre-mRNA splicing

Therapeutic potential of antisense oligonucleotides as modulators of alternative splicing

U7 snRNA-mediated correction of aberrant splicing caused by activation of cryptic splice sites

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