Stable and inheritable changes in genotype and phenotype of albino melanocytes induced by an RNA-DNA oligonucleotide View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

1998-12

AUTHORS

Vitali Alexeev, Kyonggeun Yoon

ABSTRACT

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. All black-pigmented clones exhibited a correction of the point mutation in a single allele of the tyrosinase gene. A full-length tyrosinase was detected by an antityrosinase antibody, and the enzymatic activity was restored in all converted black-pigmented clones. Only degraded fragments were detected in albino cells due to proteolytic cleavage of mutant tyrosinase. The phenotype and genotype of converted black-pigmented clones was stable. These results demonstrate a permanent and stable gene correction by the RNA-DNA oligonucleotide at the level of genomic sequence, protein, and phenotypic change by clonal analysis. More... »

PAGES

1343-1346

Journal

TITLE

Nature Biotechnology

ISSUE

13

VOLUME

16

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/4322

    DOI

    http://dx.doi.org/10.1038/4322

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1020567316

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/9853616


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