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Isolation of deletion alleles by G4 DNA-induced mutagenesis


Metazoan genomes contain thousands of sequence tracts that match the guanine-quadruplex (G4) DNA signature G3NxG3NxG3NxG3, a motif that is intrinsically mutagenic, probably because it can form secondary structures during DNA replication. Here we show how and to what extent this feature can be used to generate deletion alleles of many Caenorhabditis elegans genes.

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Figure 1: Mutagenic potential of G4 DNA tracts.
Figure 2: Sib-selection strategy to isolate deletion alleles.
Figure 3: Schematic illustration of 11 ORF-disrupting C. elegans deletion alleles that were generated by G4 DNA–mediated mutagenesis.

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We thank members of the Tijsterman lab for discussions, R. Plasterk for generous support and WormBase curators for building the platform that allows C. elegans genomic research. This work was funded by a Horizon grant from the Netherlands Genomics Initiative and a Vidi grant from ZonMw to M.T.

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D.B.P., E.K. and M.T. performed the experiments and analyzed the data. V.G. performed the bioinformatics analysis. M.T. conceived and supervised the project, and wrote the paper.

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Correspondence to Marcel Tijsterman.

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Supplementary Text and Figures

Supplementary Figures 1–2 and Supplementary Table 1 (PDF 528 kb)

Supplementary Table 2

C. elegans endogenous G4- sites (XLS 475 kb)

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Pontier, D., Kruisselbrink, E., Guryev, V. et al. Isolation of deletion alleles by G4 DNA-induced mutagenesis. Nat Methods 6, 655–657 (2009).

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