Recombinant DNA technique and sickle cell anemia research
KR Thomas, MR Capecchi - Science, 1997 - science.org
KR Thomas, MR Capecchi
Science, 1997•science.orgThe data presented in the report by Allyson Cole-Strauss et al.(6 Sept., p. 1386) infer a
remarkable phenomenon: 50 to 80% of mutant, β S-globin loci in a population of B cells
were converted to wild-type alleles after exposure of those cells to oligonucleotides
containing wild-type, β A, sequences. This represents an absolute recombination frequency
(recombinant cells/exposed cells) that is three to six orders of magnitude higher than that
normally seen in cultured mammalian cells (1).These data were published without the clonal …
remarkable phenomenon: 50 to 80% of mutant, β S-globin loci in a population of B cells
were converted to wild-type alleles after exposure of those cells to oligonucleotides
containing wild-type, β A, sequences. This represents an absolute recombination frequency
(recombinant cells/exposed cells) that is three to six orders of magnitude higher than that
normally seen in cultured mammalian cells (1).These data were published without the clonal …
The data presented in the report by Allyson Cole-Strauss et al.(6 Sept., p. 1386) infer a remarkable phenomenon: 50 to 80% of mutant, β S-globin loci in a population of B cells were converted to wild-type alleles after exposure of those cells to oligonucleotides containing wild-type, β A, sequences. This represents an absolute recombination frequency (recombinant cells/exposed cells) that is three to six orders of magnitude higher than that normally seen in cultured mammalian cells (1).
These data were published without the clonal isolation of a single recombinant cell line. Gene conversion was assayed on pooled-cell extracts containing a mixture of reagent oligonucleotides and chromosomal DNA in which the wild-type/mutant sequence ratio approached 10 8/1. Under such conditions, the potential for assay artifact should be considered, yet neither a zero time point nor an end point, in the form of cloned cells, was performed.
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