The development and regulation of gene repair

Liu L, Parekh-Olmedo H, Kmiec EB Nat. Rev. Genet. 2003 Sep;4(9):679-89 PMID: 12951569 Abstract A technique that can direct the repair of a genetic mutation in a human chromosome using the DNA repair machinery of the cell is under development. Although this approach is not as mature as other forms of gene therapy and fundamental

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Targeted nucleotide repair of cyc1 mutations in Saccharomyces cerevisiae directed by modified single-stranded DNA oligonucleotides

Brachman EE, Kmiec EB Genetics 2003 Feb;163(2):527-38 PMID: 12618392 Abstract Modified single-stranded DNA oligonucleotides have been used to direct base changes in the CYC1 gene of Saccharomyces cerevisiae. In this process, the oligonucleotide is believed to hybridize to the target site through the action of a DNA recombinase and, once bound, DNA repair enzymes act

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CRISPR/Cas9-Directed Reassignment of the GATA1 Initiation Codon in K562 Cells to Recapitulate AML in Down Syndrome

Bloh KM, Bialk PA, Gopalakrishnapillai A, Kolb EA, Kmiec EB Mol Ther Nucleic Acids 2017 Jun;7:288-298 PMID: 28624204 Abstract Using a CRISPR/Cas9 system, we have reengineered a translational start site in the GATA1 gene in K562 cells. This mutation accounts largely for the onset of myeloid leukemia in Down syndrome (ML-DS). For this reengineering, we

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Insertional Mutagenesis by CRISPR/Cas9 Ribonucleoprotein Gene Editing in Cells Targeted for Point Mutation Repair Directed by Short Single-Stranded DNA Oligonucleotides

Rivera-Torres N, Banas K, Bialk P, Bloh KM, Kmiec EB PLoS ONE 2017;12(1):e0169350 PMID: 28052104 Abstract CRISPR/Cas9 and single-stranded DNA oligonucleotides (ssODNs) have been used to direct the repair of a single base mutation in human genes. Here, we examine a method designed to increase the precision of RNA guided genome editing in human cells

Continue reading Insertional Mutagenesis by CRISPR/Cas9 Ribonucleoprotein Gene Editing in Cells Targeted for Point Mutation Repair Directed by Short Single-Stranded DNA Oligonucleotides

Analyses of point mutation repair and allelic heterogeneity generated by CRISPR/Cas9 and single-stranded DNA oligonucleotides

Bialk P, Sansbury B, Rivera-Torres N, Bloh K, Man D, Kmiec EB Sci Rep 2016 Sep;6:32681 PMID: 27609304 Abstract The repair of a point mutation can be facilitated by combined activity of a single-stranded oligonucleotide and a CRISPR/Cas9 system. While the mechanism of action of combinatorial gene editing remains to be elucidated, the regulatory circuitry

Continue reading Analyses of point mutation repair and allelic heterogeneity generated by CRISPR/Cas9 and single-stranded DNA oligonucleotides

Genetic spell-checking: gene editing using single-stranded DNA oligonucleotides

Rivera-Torres N, Kmiec EB Plant Biotechnol. J. 2016 Feb;14(2):463-70 PMID: 26402400 Abstract Single-stranded oligonucleotides (ssODNs) can be used to direct the exchange of a single nucleotide or the repair of a single base within the coding region of a gene in a process that is known, generically, as gene editing. These molecules are composed of

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Regulation of Gene Editing Activity Directed by Single-Stranded Oligonucleotides and CRISPR/Cas9 Systems

Bialk P, Rivera-Torres N, Strouse B, Kmiec EB PLoS ONE 2015;10(6):e0129308 PMID: 26053390 Abstract Single-stranded DNA oligonucleotides (ssODNs) can direct the repair of a single base mutation in human genes. While the regulation of this gene editing reaction has been partially elucidated, the low frequency with which repair occurs has hampered development toward clinical application.

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Is the age of genetic surgery finally upon us?

Kmiec EB Surg Oncol 2015 Jun;24(2):95-9 PMID: 25936245 Abstract This review discusses gene editing and its potential in oncology. Gene editing has not evolved faster towards clinical application because of its difficulty in implementation. There have been many limitations of the tools thought to be useful in therapeutic gene editing. However, recently the combinatorial use

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