Targeted nucleotide exchange in the alkaline phosphatase gene of HuH-7 cells mediated by a chimeric RNA/DNA oligonucleotide

Kren, Betsy T.; Cole-Strauss, Allyson; Kmiec, Eric B.; Steer, Clifford J.

doi:10.1002/hep.510250626

Cited by 125 publications

(102 citation statements)

References 33 publications

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“…135 Chimeraplasty was originally performed on episomal targets, 136 the sickle cell mutation in cultured lymphoblasts, 137 cell-free extracts, 138 and a variety of cultured cells, 139,140 including HuH-7 cells for the endogenous alkaline phosphatase gene. 141 To increase hepatocyte uptake, chimeric molecules were either complexed with lactosylated PEI or encapsulated with anionic, cationic, or neutral liposomes containing galactocerebroside for targeting to the asialoglycoprotein receptor. Delivery was assessed by confocal microscopy, and single base correction rates by polymerase chain reaction (PCR)/filter lift hybridizations.…”

Section: Gene Repair Strategiesmentioning

confidence: 99%

Gene therapy as an alternative to liver transplantation

Kren

Chowdhury

et al. 2002

Liver Transplantation

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Liver transplantation has become a well-recognized therapy for hepatic failure resulting from acute or chronic liver disease. It also plays a role in the treatment of certain inborn errors of metabolism that do not directly injure the liver. In fact, the liver maintains a central role in many inherited and acquired genetic disorders. There has been a considerable effort to develop new and more effective gene therapy approaches, in part, to overcome the need for transplantation as well as the shortage of donor livers. Traditional gene therapy involves the delivery of a piece of DNA to replace the faulty gene. More recently, there has been a growing interest in the use of gene repair to correct certain genetic defects. In fact, targeted gene repair has many advantages over conventional replacement strategies. In this review, we will describe a variety of viral and nonviral strategies that are now available to the liver. The ever-growing list includes viral vectors, antisense and ribozyme technology, and the Sleeping Beauty transposon system. In addition, targeted gene repair with RNA/DNA oligonucleotides, small-fragment homologous replacement, and triplex-forming and single-stranded oligonucleotides is a long-awaited and potentially exciting approach. Although each method uses different mechanisms for gene repair and therapy, they all share a basic requirement for the efficient delivery of DNA. (Liver

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Section: Gene Repair Strategiesmentioning

confidence: 99%

Gene therapy as an alternative to liver transplantation

Kren

Chowdhury

et al. 2002

Liver Transplantation

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“…This study provided the basis for subsequent applications of RDO for conversion of chromosomal genes. Since then, site-specific chromosomal corrections or mutations have been reported in several genes (13)(14)(15)(16)(17)(18)(19)(20)(21). An RDO was shown to induce a sequence-specific mutation in the factor IX gene in cell culture and in vivo by intravenous injection (19).…”

Section: Application Of Rdo In Mammalian Cellsmentioning

confidence: 99%

“…These results demonstrated the generality for a targeted sequence in vitro and in vivo by a double-stranded RDO. However, most previous studies detected the gene conversion event by PCR amplification of genomic DNA without direct confirmation at the protein level (13)(14)(15)(16). Concerns have been raised about potential artifacts during PCR amplification, or by cross-contamination between cells, especially because no clonal analysis has been performed (33,34).…”

Section: Application Of Rdo In Mammalian Cellsmentioning

confidence: 99%

“…An experimental strategy, capable of site-specific correction of single-base mutations of a target sequence, has been developed using an RNA-DNA hybrid oligonucleotide (12). The RNA-DNA oligonucleotide (RDO) was shown to either correct or cause a specific point mutation in episomal and genomic DNA in mammalian cells, in plants and in animals (12)(13)(14)(15)(16)(17)(18)(19)(20)(21). RNA has been shown facilitate DNA recombination.…”

Section: Introductionmentioning

confidence: 99%

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Gene Correction by RNA‐DNA Oligonucleotides

Alexeev

Yoon

2000

Pigment Cell Research

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of several hairs and DOPA staining of hair follicles in the An oligonucleotide composed of a contiguous stretch of RNA and DNA residues has been developed to facilitate the correctreated skin of albino mice. Such RDOs might hold a promise tion of single-base mutations of episomal and chromosomal as a therapeutic method for the treatment of skin diseases. targets in mammalian cells. The design of the oligonucleotide However, the frequency of gene correction varies among different cells, indicating that cellular activities, such as reexploited the highly recombinogenic RNA-DNA hybrids and featured hairpin capped ends avoiding destruction by cellular combination and repair, may be important for gene conversion helicases or exonucleases. The RNA-DNA oligonucleotide by RDOs. As this technology becomes more widely utilized in the scientific community, it will be important to understand (RDO) was designed to correct a point mutation in the the mechanism and to optimize the design of RDOs to imtyrosinase gene and caused a permanent gene correction in mouse albino melanocytes, determined by clonal analysis at prove their efficiency and general applicability. the level of genomic sequence, protein and phenotypic change. Recently, we demonstrated correction of the tyrosinase gene

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“…9,10 This novel approach of gene repair, based on in vitro experimental evidence, was shown by Yoon et al 11 and Cole-Strauss et al 12 to be capable of introducing targeted single base conversions in episomal and genomic DNA in cultured cells. We then reported that the chimeraplast technology could introduce a missense mutation in genomic DNA in human hepatoma cells, 13 and nonreplicating isolated rat hepatocytes. 14 The high rates of nucleotide conversion observed in primary hepatocytes resulted, in part, from significantly improved delivery of chimeraplasts using our modified nonviral-ASGPR receptor targeted delivery systems.…”

mentioning

confidence: 99%

Modification of hepatic genomic DNA using RNA/DNA oligonucleotides

Kren¹,

Chen

Felsheim

et al. 2002

Gene Ther

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Inherited metabolic diseases impose a significant worldwide burden both financially and socially. Although there have been advances in treating these disorders, alternative therapeutic approaches are being developed to correct the responsible genetic defects. Gene augmentation using viral vectors and transgene expression to replace an absent or defective protein has shown limited success in treating a variety of genetic disorders. In fact, the mechanisms by which cells and organisms protect against viral infections have presented significant barriers for long-term stable expression of the introduced transgene. In part, due to the disadvantages of using biological vectors, a variety of different approaches for nonviral gene delivery have been developed. 1 In particular, targeting of nonviral delivery systems to specific tissues by receptor/ligand interactions has been exploited for in vivo gene transfer. 2 By targeting the asialoglycoprotein receptor (ASGPR), gene delivery to hepatocytes has been significantly improved both in vitro 3 and in vivo. 4,5 Thus, we have focused on using nonviral approaches to correct the genetic defect in hepatocytes using galactosylated compounds attached directly to polycations, or incorporated in liposomes. 6 The strategy of in situ genomic repair using chimeric RNA/DNA molecules (chimeraplasts) developed from studies investigating molecular aspects of DNA repair. It became apparent that there was a significant increase in pairing efficiency between an oligonucleotide Յ50 bases and a genomic DNA target if RNA replaced DNA in a portion of the targeting oligonucleotide. 7,8 The original chimeric 68-mer design incorporated 10 2'-O-methylated RNA residues flanking each side of a 5 bp stretch of DNA, poly-T hairpin loops, a 3' GC clamp and a complementary all-DNA strand resulting in a stable, nucleaseresistant duplex molecule. The 25 bp homology segment between the chimeraplast and its genomic target is constructed such that it includes one mismatch with the DNA sequence of the gene. This engineered mismatch permits the directed alteration of the genomic target using the chimeraplast as a template for the intended correction of target sequence. It is postulated that the 'mismatched' chimeraplast complexes with the genomic DNA and creates the illusion of a genetic mutation leading to the recruitment of endogenous repair functions. 9,10 This novel approach of gene repair, based on in vitro experimental evidence, was shown by Yoon et al 11 and Cole-Strauss et al 12 to be capable of introducing targeted single base conversions in episomal and genomic DNA in cultured cells. We then reported that the chimeraplast

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Targeted nucleotide exchange in the alkaline phosphatase gene of HuH-7 cells mediated by a chimeric RNA/DNA oligonucleotide

Cited by 125 publications

References 33 publications

Gene therapy as an alternative to liver transplantation

Gene therapy as an alternative to liver transplantation

Gene Correction by RNA‐DNA Oligonucleotides

Modification of hepatic genomic DNA using RNA/DNA oligonucleotides

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