Biochips,preparation and uses

• US 20030059809A1
• Filed: 10/22/2002
• Published: 03/27/2003
• Est. Priority Date: 12/17/1999
• Status: Abandoned Application

• Current Assignee: Centre National De La Recherche Scientifique
• Original Assignee: Centre National De La Recherche Scientifique
• Inventors: Mallet, Jacques, Vujasinovic, Todor, Dumas, Sylvie
• Application Number:
  US10/168,143
• Publication Number:
  US 20030059809A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence

Abstract

The present invention concerns micro-arrays, their preparation and their uses. In particular, it concerns micro-arrays composed of nucleic acids immobilised on a support by means of arms in arborescent form and/or arms carrying a negative charge and/or directly on the supports carrying a negative charge. The methods and micro-arrays according to the present invention can be used for genetic expression detection or analysis, for research of genes of interest, or for diagnostic applications, for example.

Claims

1. Micro-array characterised in that it comprises nucleic acids immobilised on a support by means of arborescent shaped arms, comprising a polymer of biological origin.

2. Micro-array according to claim 1, characterised in that the arm is a branched polymer with one or several branching levels.

3. Micro-array according to claim 1 or 2, characterised in that the arm is an organic polymer.

4. Micro-array according to any one of claims 1 to 3, characterised in that the arm is a sugar polymer, for example glycogen or a glycogen derivative or amylopectin.

5. Micro-array according to any one of claims 1 to 3, characterised in that the arm is a branched polymer comprising repeated galactose, glucose, mannose, fucose, xylose, N-acetylgalactosamine and/or N-acetylglucosamine monomers.

6. Micro-array according to any one of claims 1 to 3, characterised in that the arm is a glycopolypeptide, for example aggrecane.

7. Micro-array according to any one of claims 1 to 3, characterised in that the arm is a polypeptide compound, for example an immunoglobulin.

8. Micro-array according to any one of the preceding claims, characterised in that the nucleic acids are fixed in covalent mode to the ends of the arborescent arms.

9. Micro-array according to any one of the preceding claims, characterised in that the arms are fixed in covalent mode to the support by the trunk part of the molecule.

10. Micro-array according to any one of the preceding claims, characterised in that the support has a flat and/or convex surface.

11. Micro-array according to claim 10, characterised in that the support is a solid or semi-solid support.

12. Micro-array according to claim 11, characterised in that the support is composed of glass, silica, poly-lysine, amino-silanes and/or amino-reactive silanes.

13. Micro-array according to any one of the preceding claims, characterised in that the nucleic acids are single or double strand nucleic acids, of natural, synthetic and/or semi-synthetic origin.

14. Micro-array according to claim 13, characterised in that the nucleic acids are chosen among synthesized oligonucleotides, PCR products, genes or gene fragments, plasmids, single or double strand cDNA or RNA.

15. Micro-array according to claim 14, characterised in that the nucleic acids are single strand nucleic acids, notably molecules of single strand nucleic acids with a length between approximately 25 and 100 nucleotides, preferably between approximately 30 and 60 nucleotides.

17. Micro-array according to claim 13, characterised in that the nucleic acids are single strand nucleic acids with a length of approximately 30 to 60 nucleotides.

18. Micro-array according to claim 17, characterised in that the single strand nucleic acids are single strand DNAs with a length of approximately 30 to 60 nucleotides.

19. Micro-array according to claim 17, characterised in that the single strand nucleic acids are single strand RNAs with a length of approximately 30 to 60 nucleotides.

20. Micro-array according to any one of claims 17 to 19, characterised in that the single strand nucleic acids are nucleic acids produced by chemical synthesis in vitro.

21. Micro-array according to any one of claims 17 to 20, characterised in that it comprises RNAs immobilised on a solid or semi-solid support by means of arborescent form arms comprising a polymer of biological origin.

22. Micro-array according to any one of claims 17 to 20, characterised in that it comprises nucleic acids produced by PCR immobilised on a solid or semi-solid support by means of arborescent form arms comprising a polymer of biological origin.

23. Micro-array according to any one of claims 1 to 15 and 17 to 22, characterised in that the arm carries a negative electrical charge.

24. Micro-array according to any one of claims 1 to 15 and 17 to 22, characterised in that the support carries a uniform negative electrical charge.

25. Use of a micro-array according to any one of claims 1 to 15 and 17 to 24, for studying the regulation of genetic expression.

26. Use of a micro-array according to any one of claims 1 to 15 and 17 to 24, for research of genes or gene fragments.

27. Use of a micro-array according to any one of claims 1 to 15 and 17 to 24, for targets identification.

28. Use of a micro-array according to any one of claims 1 to 15 and 17 to 24, for genetic diagnosis.

29. Process for micro-array preparation according to any one of claims 1 to 15 and 17 to 24, characterised in that it comprises:

a) a step for fixing the arborescent form arm on the support, and

b) a step for fixing the target molecule to the arm in arborescent form obtained during a).

30. Process for micro-array preparation according to any one of claims 1 to 15 and 17 to 24, characterised in that it comprises:

a) a step for fixing the target molecule to the arm in arborescent form,

b) a step for fixing the complex obtained during a) on the support.

16. Use of a polymer of biological origin (notably a sugar polymer, and in particular derived from glycogen) with a spatial organisation in arborescent form, for fixing nucleic acids on supports.