Mutation detection using ENDO1: Application to disease diagnostics in humans and TILLING and Eco-TILLING in plants

Karine Triques, Elodie Piednoir, Marion Dalmais, Julien Schmidt, Christine Le Signor, Mark E Sharkey, Michel Caboche, Bénédicte Sturbois, Abdelhafid Bendahmane

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Background: Most enzymatic mutation detection methods are based on the cleavage of heteroduplex DNA by a mismatch-specific endonuclease at mismatch sites and the analysis of the digestion product on a DNA sequencer. Important limitations of these methods are the availability of a mismatch-specific endonuclease, their sensitivity in detecting one allele in pool of DNA, the cost of the analysis and the ease by which the technique could be implemented in a standard molecular biology laboratory. Results: The co-agroinfiltration of ENDO1 and p19 constructs into N. benthamiana leaves allowed high level of transient expression of a mismatch-specific and sensitive endonuclease, ENDO1 from Arabidopsis thaliana. We demonstrate the broad range of uses of the produced enzyme in detection of mutations. In human, we report the diagnosis of the G1691A mutation in Leiden factor-V gene associated with venous thrombosis and the fingerprinting of HIV-1 quasispecies in patients subjected to antiretroviral treatments. In plants, we report the use of ENDO1 system for detection of mutant alleles of Retinoblastoma-related gene by TILLING in Pisum sativum and discovery of natural sequence variations by Eco-TILLING in Arabidopsis thaliana. Conclusion: We introduce a cost-effective tool based on a simplified purification protocol of a mismatch-specific and sensitive endonuclease, ENDO1. Especially, we report the successful applications of ENDO1 in mutation diagnostics in humans, fingerprinting of complex population of viruses, and in TILLING and Eco-TILLING in plants.

Original languageEnglish
Article number42
JournalBMC Molecular Biology
Volume9
DOIs
StatePublished - Apr 23 2008
Externally publishedYes

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Endonucleases
Mutation
Arabidopsis
Genes
Nucleic Acid Heteroduplexes
Alleles
Retinoblastoma Genes
Costs and Cost Analysis
Molecular biology
DNA
Peas
Viruses
Venous Thrombosis
Purification
HIV-1
Costs
Molecular Biology
Digestion
Availability
Enzymes

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

Cite this

Mutation detection using ENDO1 : Application to disease diagnostics in humans and TILLING and Eco-TILLING in plants. / Triques, Karine; Piednoir, Elodie; Dalmais, Marion; Schmidt, Julien; Le Signor, Christine; Sharkey, Mark E; Caboche, Michel; Sturbois, Bénédicte; Bendahmane, Abdelhafid.

In: BMC Molecular Biology, Vol. 9, 42, 23.04.2008.

Research output: Contribution to journalArticle

Triques, K, Piednoir, E, Dalmais, M, Schmidt, J, Le Signor, C, Sharkey, ME, Caboche, M, Sturbois, B & Bendahmane, A 2008, 'Mutation detection using ENDO1: Application to disease diagnostics in humans and TILLING and Eco-TILLING in plants', BMC Molecular Biology, vol. 9, 42. https://doi.org/10.1186/1471-2199-9-42
Triques, Karine ; Piednoir, Elodie ; Dalmais, Marion ; Schmidt, Julien ; Le Signor, Christine ; Sharkey, Mark E ; Caboche, Michel ; Sturbois, Bénédicte ; Bendahmane, Abdelhafid. / Mutation detection using ENDO1 : Application to disease diagnostics in humans and TILLING and Eco-TILLING in plants. In: BMC Molecular Biology. 2008 ; Vol. 9.
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