TARGETED GENE THERAPY
Subject:TGR-Non Disease Specific
(The following material is presented in Ascending Date Order)
This post is generated by a program that reads a database and generates this file. Errors will result from incorrect database material. Updates to the database automatically reflected in this file.
Researcher Name shown is my estimate of Principal Researcher
DB Key:1996-INV-A
Doc:Abstract
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Failed or Not Applicable
Of Note:Changing a glutamine or lysine codon into a stop codon through a chimeric directed DNA repair system.
2/1/1996-----Kmiec EB-----Thomas Jefferson University
Title:Genomic targeting and genetic conversion in cancer therapy
Link:http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=8607028&dopt=Abstract
Snippet:
We have developed an experimental strategy that centers around site-specific DNA base mutation or correction using a unique chimeric oligonucleotide. This chimeric molecule has demonstrated higher recombinogenic activities than identical oligonucleotides containing only DNA residues, both in vitro and in vivo. The chimeric molecule is designed to hybridize to a target site within the genome and induce a single base mismatch at the residue targeted for mutation
DB Key:1996-INV-B
Doc:Abstract
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Failed or Not Applicable
Of Note:First announcement of Targeted Gene Repair
3/1/1996-----Kmiec EB-----Thomas Jefferson University
Title:Targeted gene correction of episomal DNA in mammalian cells mediated by a chimeric RNA.DNA oligonucleotide
Link:http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=8700887&dopt=Abstract
Snippet:
An experimental strategy to facilitate correction of single-base mutations of episomal targets in mammalian cells has been developed. The method utilizes a chimeric oligonucleotide composed of a contiguous stretch of RNA and DNA residues in a duplex conformation with double hairpin caps on the ends. The RNA/DNA sequence is designed to align with the sequence of the mutant locus and to contain the desired nucleotide change. Activity of the chimeric molecule in targeted correction was tested in a model system in which the aim was to correct a point mutation in the gene encoding the human liver/bone/kidney alkaline phosphatase
DB Key:1998-INV-A
Doc:Web Page Article
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Successful
Of Note:Introduced a mutation in the Hemophilia B gene in rodents
3/5/1998-----Steer CJ-----University of Minnesota
Title:Researchers perform gene surgery 'Chimeraplasty' for first time in medicine; reported March issue Nature Medicine
Link:http://hdlighthouse.org/see/genetherapy/chimeraplasty.htm
Snippet:
Kimeragen, Inc. today announced that researchers at the University of Minnesota (Minneapolis) have utilized the Company's novel patented gene alteration, repair and correction technology, termed chimeraplasty, to introduce a mutation in the Hemophilia B gene in rodents. The results of this study were reported in the March issue of “Nature Medicine.” This study demonstrates for the first time in medical history that alteration of genes within live animals can be efficiently accomplished by intravenous injection of a synthetic drug that targets and changes a pre-selected section of DNA. According to the Minneapolis researchers, the DNA repair technique, chimeraplasty, should be applicable to many other inherited or acquired diseases.
Kimeragen, Inc. today announced that researchers at the University of Minnesota (Minneapolis) have utilized the Company's novel patented gene alteration, repair and correction technology, termed chimeraplasty, to introduce a mutation in the Hemophilia B gene in rodents. The results of this study were reported in the March issue of “Nature Medicine.” This study demonstrates for the first time in medical history that alteration of genes within live animals can be efficiently accomplished by intravenous injection of a synthetic drug that targets and changes a pre-selected section of DNA. According to the Minneapolis researchers, the DNA repair technique, chimeraplasty, should be applicable to many other inherited or acquired diseases.
DB Key:1998-INV-C
Doc:Abstract
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Failed or Not Applicable
Of Note:One end of the double hairpin may unwind to form a 14-base-RecA filament which initiates the reaction.
6/1/1998-----Kmiec EB-----Thomas Jefferson University
Title:RecA-mediated joint molecule formation between O-methylated RNA/DNA hairpins and single-stranded targets
Link:http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9671026&dopt=Abstract
Snippet:
Chimeric oligonucleotides consisting of one DNA strand paired with an O-methylated RNA strand interrupted by six DNA residues have been used in gene targeting experiments. Here we demonstrate that these hairpins can form a heteroduplex (or joint molecule) with single-stranded DNA targets in a reaction mediated by the E. coli RecA protein. One end of the double hairpin may unwind to form a 14-base-RecA filament which initiates the reaction. Chimeric oligonucleotides containing only O-methylated RNA residues on one strand or truncated hairpins lacking this 14-base segment did not participate in RecA-driven heteroduplex formation under these reaction conditions. The results presented here represent a first step in studying one facet of a strategy which uses O-methylated RNA residues as participants in homologous pairing events
DB Key:1998-INV-D
Doc:Abstract
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Failed or Not Applicable
Of Note:Kimeragen Inc
10/4/1998-----Kmiec EB-----Kimeragen, Inc
Title:Targeted gene correction: a new strategy for molecular medicine
Link:http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9793931&dopt=Abstract
Snippet:
A number of strategies are presently being used to replace or augment a dysfunctional gene with a correct copy of itself. Now, a novel approach to correct the dysfunctional gene in the chromosome is being developed. Data obtained from biochemical, cell-based and animal studies suggest that the era of gene repair is dawning. It is now conceivable that inherited and non-inherited disorders might be treated with a small molecular tool designed to fix the mutation directly. Here, the conceptualization of the technique and its barriers to success are discussed
DB Key:1998-INV-E
Doc:Abstract
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Successful
Of Note:First pigment change
12/1/1998-----Yoon, Kyonggeun-----Thomas Jefferson University
Title:Stable and inheritable changes in genotype and phenotype of albino melanocytes induced by an RNA-DNA oligonucleotide
Link:http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9853616&dopt=Abstract
Snippet:
Experimental strategies have been developed to correct point mutations using chimeric oligonucleotides composed of RNA and DNA. We used these RNA-DNA oligonucleotides to correct a point mutation in mouse tyrosinase, a key enzyme for melanin synthesis and pigmentation. Melanocytes derived from albino mice contain a homozygous point mutation (TGT-->TCT) in the tyrosinase gene, resulting in an amino acid change from Cys-->Ser. Correction of this point mutation results in the restoration of tyrosinase activity and melanin synthesis, thus changing the pigmentation of the cells. Upon transfection of the RNA-DNA oligonucleotide to albino melanocytes, we detected black-pigmented cells and isolated multiple single clones
DB Key:1999-INV-B
Doc:Web Page Article
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Failed or Not Applicable
Of Note:Types of Gene Vectors
1/1/1999-----Baxter Health Care-----Baxter Health Care
Title:Types of Vectors
Link:http://www.hemophiliagalaxy.com/1_PATIENTS/Q_A/future/directory_topics/SEC_03d/
Snippet:
Describes the 7 Vector Types
DB Key:1999-INV-C
Doc:Abstract
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Failed or Not Applicable
Of Note:Can be corrected at frequencies of approximately 0.1% and 0.005%, respectively
3/1/1999-----Kmiec EB-----Thomas Jefferson University
Title:Targeted gene repair directed by the chimeric RNA/DNA oligonucleotide in a mammalian cell-free extract
Link:http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9973621&dopt=Abstract
2nd Link:http://nar.oupjournals.org/cgi/content/abstract/27/5/1323
Snippet:
Using a mammalian cell-free extract and a genetic readout in Escherichia coli, we find that point mutations and single base deletions can be corrected at frequencies of approximately 0.1% and 0.005%, respectively. The reaction depends on an accurately designed chimera and the presence of functional hMSH2 protein. The results of genetic and biochemical studies reported herein suggest that the process of mismatch repair functions in site-directed gene correction.
DB Key:1999-INV-D
Doc:Abstract
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Failed or Not Applicable
Of Note:The chimeric molecules have been demonstrated to be effective in the alteration of single nucleotides in episomal and genomic DNA in cell culture, as well as genomic DNA of cells in situ.
4/1/1999-----Steer CJ-----University of Minnesota
Title:Gene repair using chimeric RNA/DNA oligonucleotides
Link:http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10349687&dopt=Abstract
Snippet:
The chimeric molecules are designed with a homologous targeting sequence comprised of a DNA region flanked by blocks of 2'-O-methyl RNA residues (the chimeric strand), its complementary all-DNA strand, thymidine hairpin caps, a single-strand break, and a double-stranded clamp region. The oligonucleotide can align in perfect register with a genomic target except for the designed single base pair mismatch, which is recognized and corrected by harnessing the cell's endogenous DNA repair system
DB Key:1999-INV-G
Doc:Abstract
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Failed or Not Applicable
Of Note:None
10/1/1999-----Lanzov VA-----Russian Academy of Sciences
Title:Gene targeting for gene therapy: prospects
Link:http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10527679&dopt=Abstract
Snippet:
Based on the model of homologous recombination for the well-studied prokaryotic and the less studied eukaryotic systems, three approaches have been employed to improve the efficiency and accuracy of homologous recombination events. These are: (1) artificial double-strand breaks in both the exogenous and the chromosomal DNA, (2) a contiguous long homology between the exogenous and chromosomal DNA, and (3) a transient overproduction of an active recombinase, the bacterial RecA or mammalian RecA-like proteins, in mammalian cell nuclei.
DB Key:1999-INV-I
Doc:European Society of Gene Therapy Seventh Meeting
European Society of Gene Therapy Seventh Meeting
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Failed or Not Applicable
Of Note:Blaese M (Kimeragen – PA – USA) described an alternative approach to gene correction through the use of chimeric oligonucleotides consisting of both DNA and RNA, called chimeraplasts
11/1/1999-----Blaese, Michael R. M.D.-----Valigen
Title:Genetic and other organ diseases
Link:http://www.fisv.org/report_ciliberto.html
Snippet:
Structurally, chimeraplasts consist of a stretch of 2'O-methyl RNA homologous to the target sequence except for a single mismatch located at the center of the sequence. The complementary strand is made of deoxyribonucleotides. Two DNA loops flank the double strand structure. A nick placed at the 5' end of the RNA strand interrupts the integrity of the entire structure. These chimeric oligonucleotides have been shown to introduce specific changes in the genomic DNA of bacteria carrying a plasmid with a single inactivating mutation in the Kan-res. gene.
DB Key:1999-INV-L
Doc:Abstract
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Failed or Not Applicable
Of Note:The variability in gene conversion frequency by an RNA-DNA oligonucleotide (RDO) prompted us to develop a system as a means of measuring the conversion frequency rapidly and reproducibly
12/1/1999-----Yoon, Kyonggeun-----Thomas Jefferson University
Title:A sequence-specific gene correction by an RNA-DNA oligonucleotide in mammalian cells characterized by transfection and nuclear extract using a lacZ shuttle system
Link:http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10637447&dopt=Abstract
Snippet:
Nuclear extracts from DT40 cells, which have a higher homologous recombination rate than any other mammalian cells exhibited 0.1-0.6% of gene correction. These results indicated that recombination may be rate-limiting in gene conversion by RDO in cells with competent mismatch repair activities.
DB Key:2000-INV-A
Doc:Abstract
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Successful
Of Note:First pigment change
1/1/2000-----Yoon, Kyonggeun-----Thomas Jefferson University
Title:Localized in vivo genotypic and phenotypic correction of the albino mutation in skin by RNA-DNA oligonucleotide
Link:http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=10625389
Snippet:
We recently demonstrated that an RNA-DNA oligonucleotide corrected a point mutation in the mouse tyrosinase gene, resulting in permanent and inheritable restoration of tyrosinase enzymatic activity, melanin synthesis, and pigmentation changes in cultured melanocytes. In this study, we extended gene correction of melanocytes from tissue culture to live animals, using a chimeric oligonucleotide designed to correct a point mutation in the tyrosinase gene. Both topical application and intradermal injection of this oligonucleotide to albino BALB/c mouse skin resulted in dark pigmentation of several hairs in a localized area
DB Key:2000-INV-A1B
Doc:Web Page Article
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Successful
Of Note:First pigment change
1/10/2000-----Web Article-----Web Page
Title:Gene Repair Adds Pigment To Hair
Link:http://www.applesforhealth.com/addpigment1.html
Snippet:
Web Article
DB Key:2000-INV-A1A
Doc:Lycos Health
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Failed or Not Applicable
Of Note:First pigment change
1/10/2000-----Web Article-----Web Page
Title:Experts Say Technique Could Provide 'Factory' for Gene Therapy
Link:http://webmd.lycos.com/content/article/1728.54014
Snippet:
Web Article
DB Key:2000-INV-B
Doc:Abstract
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Failed or Not Applicable
Of Note:We used a fluorescent labelled pure 68-mer DNA-analogue of a chimeric oligonucleotides to follow the intracellular fate of these kind of genetic material
3/10/2000-----Welz C-----Philipps University
Title:Nuclear gene targeting using negatively charged liposomes
Link:http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10699729&dopt=Abstract
Snippet:
The AVE-3 formulation shows enhanced properties compared to a classical neutral and negatively charged formulation. Nuclear localisation of oligos could only be observed with the AVE-3 formulation. Furthermore only the negatively charged liposome formulations interact with the protamine-complexed oligonucleotides
DB Key:2000-INV-C
Doc:Abstract
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Failed or Not Applicable
Of Note:The design of the oligonucleotide exploited the highly recombinogenic RNA-DNA hybrids and featured hairpin capped ends avoiding destruction by cellular helicases or exonucleases
4/13/2000-----Yoon, Kyonggeun-----Thomas Jefferson University
Title:Gene correction by RNA-DNA oligonucleotides
Link:http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10841028&dopt=Abstract
Snippet:
However, the frequency of gene correction varies among different cells, indicating that cellular activities, such as recombination and repair, may be important for gene conversion by RDOs.
DB Key:2000-INV-D
Doc:The World and I Online
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Failed or Not Applicable
Of Note:chimeraplasty relies on short, synthetically produced sequences of DNA/RNA hybrids (chimeraplasts) to interact with the faulty genetic material and to stimulate the cell's genetic repair mechanism to correct the defects.
5/1/2000-----Sharrer, Terry G.-----Curator of health sciences at the Smithsonian Institution in Washington
Title:The Promise of Genetic Cures
Link:http://www.worldandi.com/public/2000/may/medgene.html
Snippet:
The first clinical protocol, tested on rats, was designed to treat Crigler-Najjar syndrome--a genetic disease in which a defective liver enzyme fails to break down bilirubin, which at high levels can damage the central nervous system. The chimeraplasts were cleverly introduced into the nuclei of liver cells, where the chimeraplasts' sequence matched almost perfectly with a complementary sequence on a chromosome. But a mismatch of one nucleotide, at the point mutation linked to the disease, activated the cellular DNA repair enzymes to fix the defective gene. Widening the applications for this approach would mainly depend on finding the delivery systems for specific types of cells.
DB Key:2000-INV-E
Doc:Abstract
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Failed or Not Applicable
Of Note:By contrast, the chimeric strand of the oligonucleotide does not function as a template for gene repair
5/16/2000-----Kmiec EB-----University of Delaware
Title:A plausible mechanism for gene correction by chimeric oligonucleotides
Link:http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10801331&dopt=Abstract
Snippet:
Analysis of structure-activity relationships demonstrates that the DNA strand of the chimeric oligonucleotide acts as a template for high-fidelity gene correction when one of its bases is mismatched to the targeted gene. By contrast, the chimeric strand of the oligonucleotide does not function as a template for gene repair. Instead, it appears to augment the frequency of gene correction by facilitating complex formation with the target. In the presence of RecA protein, each strand of a chimeric oligonucleotide can hybridize with double-stranded DNA to form a complement-stabilized D-loop. This reaction, which may take place by reciprocal four-strand exchange, is not observed with oligonucleotides that lack 2'-O-methyl RNA segments. Preliminary sequencing data suggest that complement-stabilized D-loops may be weakly mutagenic. If so, a low level of random mutagenesis in the vicinity of the chimera binding site may accompany gene repair.
DB Key:2000-INV-G
Doc:Cancer Detection and Prevention 2000
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Failed or Not Applicable
Of Note:Chimeraplasty has been used successfully to change DNA sequences in bacteria, yeast, plants, rodent and human cells
10/28/2000-----Blaese, RM MD-----Valigen
Title:Gene repair therapy with chimeric RNA-DNA oligonucleotides
Link:http://www.cancerprev.org/Journal/Issues/24/101/312/3606
Snippet:
Development of improved delivery systems to other organs and cell types should enable treatment of a much wider range of inherited and acquired disorders based in the genes.
DB Key:2000-INV-H
Doc:Abstract
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Failed or Not Applicable
Of Note:Assay
11/1/2000-----Yoon, Kyonggeun-----Thomas Jefferson University
Title:Targeted single-base correction by RNA-DNA oligonucleotides
Link:http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11084691&dopt=Abstract
DB Key:2000-INV-I
Doc:Abstract
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Failed or Not Applicable
Of Note:single-stranded oligonucleotides
11/2/2000-----Kmiec EB-----University of Delaware
Title:The DNA strand of chimeric RNA/DNA oligonucleotides can direct gene repair/conversion activity in mammalian and plant cell-free extracts
Link:http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11058133&dopt=Abstract
2nd Link:http://nar.oupjournals.org/cgi/content/abstract/28/21/4332?maxtoshow=&HITS=10&hits=10&RESULTFORMAT=&fulltext=Oligonucleotide-based+gene+correction+strategies%3A+applications+to+neuromuscular+&searchid=1032108732702_810&stored_search=&FIRSTINDEX=0&sortspec=Score+desc+PUBDATE_SORTDATE+desc&fdate=1/1/1995&journalcode=nar
Snippet:
An uninterrupted stretch of DNA bases within the chimera is known to be active in the sequence alteration while RNA residues aid in complex stability. In this study, the two strands were separated in the hope of defining the role each plays in conversion. Using a series of single-stranded oligonucleotides, comprised of all RNA or DNA residues and various mixtures, several new structures have emerged as viable molecules in nucleotide conversion. When extracts from mammalian and plant cells and a genetic readout assay in bacteria are used, single-stranded oligonucleotides, containing a defined number of thioate backbone modifications, were found to be more active than the original chimera structure in the process of gene repair
DB Key:2000-INV-Q
Doc:Abstract
Vector:RDO RNA-DNA chimeric oligonucleotide
Experiment:Failed or Not Applicable
Of Note:RecA protein can catalyze a four-strand exchange
12/1/2000-----Kmiec EB-----University of Delaware
Title:Evidence for a four-strand exchange catalyzed by the RecA protein
Link:http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11106508&dopt=Abstract
Snippet:
Strand exchange between two duplexes is usually initiated as a three-strand event that requires the presence of a single-stranded overhang or gap in one of the two molecules. Here we show that the RecA protein can catalyze a four-strand exchange. Specifically, it can recombine short hairpin substrates with homologous stems provided that one of the hairpins possesses a chimeric DNA/RNA backbone. This four-strand exchange reaction goes to completion in the presence of ATPgammaS and releases a stable heteroduplex upon removal of the RecA protein |
