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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1984 Sep;81(18):5676–5679. doi: 10.1073/pnas.81.18.5676

Transient complementation of xeroderma pigmentosum cells by microinjection of poly(A)+ RNA.

R J Legerski, D B Brown, C A Peterson, D L Robberson
PMCID: PMC391773  PMID: 6207527

Abstract

An assay has been developed in which excision repair deficiency of xeroderma pigmentosum cells is transiently complemented, as measured by unscheduled DNA synthesis, by microinjection of cytoplasmic poly(A)+ RNA derived from HeLa cells. Four different complementation groups of xeroderma pigmentosum have been assayed. Groups A and G showed complementation, whereas groups D and F did not. Survival for cells in each of the groups subsequent to microinjection was approximately equal to 75%. Approximately 10-25% of surviving cells from groups A and G were complemented, as judged by near-normal unscheduled DNA synthesis. Fractionation of cytoplasmic poly(A)+ RNA on a 15-30% nondenaturing sucrose gradient and subsequent microinjection of the individual fractions indicate that repair mRNAs that complement xeroderma pigmentosum groups A and G sediment at approximately 11 S and 12 S, respectively. This assay should be of great utility in the cloning and biochemical analysis of DNA repair genes.

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Selected References

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