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Process for detection of DNA modifications and protein binding by single molecule manipulation

A molecular, protein-based technology used in the detection of DNA modifications and protein binding by single-molecule manipulation, which can solve problems such as the lack of insights into genome-wide biological functions, improve statistics, reduce instrument drift, and eliminate false positives

Active Publication Date: 2018-09-28
CENT NAT DE LA RECH SCI (C N R S) +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although many of these principles appear to be valid when studying other genes, these principles often do not provide genome-wide insights into biological function

Method used

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  • Process for detection of DNA modifications and protein binding by single molecule manipulation
  • Process for detection of DNA modifications and protein binding by single molecule manipulation
  • Process for detection of DNA modifications and protein binding by single molecule manipulation

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Embodiment Construction

[0195] Experimental example

Background of the invention

[0196] Binding of proteins to DNA is an important phenomenon in biology; it is a very general process that controls many reactions. While the thermodynamic equilibrium nature of this mechanism is well known, measuring its kinetics is a more challenging problem. The use of single molecules not only provides the ability to measure the time it takes a protein to find its DNA target but also the exact location of a binding event. We describe here a new single-molecule assay that achieves these goals.

[0197] Although this assay is broad in scope, we first demonstrate its applicability to binding of specific oligonucleotides, as well as non-specific binding of helicases to ssDNA.

[0198] Finally, we discuss the specific binding of antibodies that recognize methylated sites in DNA.

[0199] Summary

[0200] The present invention relates to novel methods for detecting a wide variety of DNA modification and DNA protei...

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Abstract

The present invention relates to a method for determining whether a protein binds to a specific DNA sequence. This method is useful in particular for identifying modifications to the DNA sequence (e.g. methylations) via the binding of proteins that specifically recognize those modifications (e.g. antibodies), but also to identify the binding sequence on DNA of a variety of proteins.

Description

[0001] This application is a divisional application of the Chinese patent application with the application number 201480017389.9, the application date is January 22, 2014, and the invention title is "Method for detecting DNA modification and protein binding through single-molecule manipulation". technical field [0002] The present invention relates to methods for determining whether a protein binds to a specific DNA sequence. This method is particularly useful for identifying DNA sequence modifications (eg, methylation) through the binding of proteins (eg, antibodies) that specifically recognize these modifications, and also for identifying binding sequences of various proteins on DNA. Background technique [0003] Binding of proteins to DNA is an important phenomenon in biology; it has fundamental roles in regulating cellular and viral functions. These include fundamental cellular processes such as DNA replication, transcription, DNA repair and DNA recombination, but also ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883
CPCG01N33/557C12Q1/6874G01N33/53G01N33/577C12Q1/6883C12Q2600/154C12Q2600/156
Inventor D·邦西蒙V·克罗凯特H·古埃J-F·阿莱曼德丁方圆
Owner CENT NAT DE LA RECH SCI (C N R S)
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