TARGETED GENE THERAPY
Subject:Hepatic Genes
(The following material is presented in Ascending Date Order)
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Researcher Name shown is my estimate of Principal Researcher
DB Key:1997-HG-A
Doc:Abstract
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Failed or Not Applicable
Of Note:Hepatic Genes
6/25/1997-----Steer CJ-----University of Minnesota
Title:Targeted nucleotide exchange in the alkaline phosphatase gene of HuH-7 cells mediated by a chimeric RNA/DNA oligonucleotide
Link:http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9185769&dopt=Abstract
Snippet:
A chimeric 2'-O-methylated-RNA/DNA oligonucleotide containing sequences complementary to 25 bases of the alkaline phosphatase gene was constructed as a duplex containing a G to A substitution at nucleotide 935. Cells were transfected with oligonucleotides for 48 hours, then harvested for DNA isolation and polymerase chain reaction (PCR) amplification of exon 6 of the alkaline phosphatase gene. Colony lifts were hybridized to 17 mer 32P-labeled oligonucleotide probes specific to the 935-G and 935-A sequences. Hybridizing colonies were grown, plasmid DNA isolated, and sequenced
DB Key:1997-HG-A
Doc:Abstract
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Failed or Not Applicable
Of Note:Hepatic Genes
6/25/1997-----Kmiec EB-----University of Minnesota
Title:Targeted nucleotide exchange in the alkaline phosphatase gene of HuH-7 cells mediated by a chimeric RNA/DNA oligonucleotide
Link:http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9185769&dopt=Abstract
Snippet:
A chimeric 2'-O-methylated-RNA/DNA oligonucleotide containing sequences complementary to 25 bases of the alkaline phosphatase gene was constructed as a duplex containing a G to A substitution at nucleotide 935. Cells were transfected with oligonucleotides for 48 hours, then harvested for DNA isolation and polymerase chain reaction (PCR) amplification of exon 6 of the alkaline phosphatase gene. Colony lifts were hybridized to 17 mer 32P-labeled oligonucleotide probes specific to the 935-G and 935-A sequences. Hybridizing colonies were grown, plasmid DNA isolated, and sequenced
DB Key:1998-HG-A
Doc:Abstract
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Failed or Not Applicable
Of Note:FACTOR IX
3/10/1998-----Steer CJ-----University of Minnesota
Title:In vivo site-directed mutagenesis of the factor IX gene by chimeric RNA/DNA oligonucleotides
Link:http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9500600&dopt=Abstract
Snippet:
Oligonucleotides were targeted to hepatocytes in cell culture or in vivo by intravenous injection. Nucleotide conversion was both site-specific and dose-dependent.
DB Key:2002-HG-M
Doc:Science Frontier
Vector:RDO/SFHR/TFO RNA-DNA chimeric oligonucleotide
Experiment:Failed or Not Applicable
Of Note:Hepatic Genes
3/1/2002-----Steer CJ-----University of Minnesota
Title:Gene Repair in the New Age
of Gene Therapy
Link:http://hepatitis-central.com/hcv/info/2002/mar/hepcentral-512-generepair.pdf
2nd Link:http://www.hepnet.com/
Snippet:
The potential of the chimeric molecule to repair genetic defects
was first shown by correction of an episomal target and then repair
of the sickle cell mutation in lymphoblasts.12,13 The unexpectedly
high levels of gene conversion prompted additional in vivo studies
to rule out potential artifacts. Delivery systems were developed
with liposomes and cationic polymers that were targeted to the
asialoglycoprotein receptor on hepatocytes. In both hepatoma cells
and primary hepatocytes there was a direct correlation between
receptor-mediated delivery, nuclear uptake, and nucleotide conversion.
14,15 Subsequently, an RNA/DNA oligonucleotide was
used in vivo to mutate the factor IX gene in hepatocytes.16 The
chimeraplast was designed to introduce a single base mutation in
the gene, rendering the factor IX protein nonfunctional. Lactosylated
polyethylenimine (PEI), a branched cationic polymer, was
used to compact the molecules and deliver them to the liver.
Greater than 40% of the factor IX gene pool was mutated and
resulted in a similar decrease in factor IX clotting activity. Most
importantly, these changes remained stable for 2 years in both
quiescent and regenerated liver after 70% partial hepatectomy.
The results of this first in vivo study signaled the dawn of gene
repair as a novel therapeutic approach to the treatment of genetic
disease.
The same strategy was then used to correct the base deletion in
the UGT1A1 gene in the Gunn rat model of Crigler-Najjar syndrome
type 1.17 The chimeraplasts were either encapsulated in
liposomes or compacted with lactosylated PEI and targeted to
hepatocytes. Filter lift hybridizations, restriction fragment length
polymorphisms (RFLP), and Southern blot analysis confirmed
that greater than 20% of the UGT1A1 gene pool in the liver wa
DB Key:2002-HG-I
Doc:Abstract
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Successful
Of Note:Hepatic Genes
6/1/2002-----Steer CJ-----University of Minnesota
Title:Modification of hepatic genomic DNA using RNA/DNA oligonucleotides
Link:http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12032688&dopt=Abstract
2nd Link:http://www.spinaweb.ie/showcase/1150/hepatocytes.htm
Snippet:
single-stranded oligonucleotides carrying a single nucleotide mismatch with the target sequence were capable of promoting gene conversion using either mitochondrial or nuclear extracts |
