You are here : Home > The lab > Luminescent probes synthesis for sequence-specific recognition of double-stranded DNA

Guillaume Frémy

Luminescent probes synthesis for sequence-specific recognition of double-stranded DNA

Published on 9 December 2019
Thesis presented December 09, 2019

DNA is the carrier of genetic information in living organisms. Targeting and visualizing in vivo a specific DNA sequence is of particular interest for medical diagnosis and biological research. In this purpose, luminescent detection is very attractive because it can be easily observed with simple tools.
The aim of this work was to establish the proof of concept of luminescent probes for the sequence-specific detection of double stranded DNA based on lanthanide luminescence, which is attractive for biological applications, and on zinc finger proteins for their DNA binding properties. We focused on the detection of a 12-base pair palindromic DNA using a pair of zinc finger proteins, one bearing a lanthanide(III) complex as a the FRET donor and the other an organic fluorophore as an acceptor.
In this purpose, a new family of bioconjugatable lanthanide(III) complexes was developed and zinc finger proteins with various chromophores were synthesized chemically, by combination of solid phase peptide synthesis and assembly of peptidic segments by native chemical ligation. The spectroscopic characterizations of these systems have evidenced the interaction of the probes with the palindromic DNA, thereby validating the proof of concept of luminescence detection of this DNA sequence by a lanthanide-based FRET system.

DNA, Zinc Fingers, Lanthanides, Native Chemical, Detection, FRET

On-line thesis.