DNA sequences comprising gene transcription regulatory qualities and methods for detecting and using such DNA sequences
AssigneeChromagenics B.V.
InventorsArie Peter Otte, Arthur Leo Kruckeberg
Life of this patent
Estimating the term…
Abstract
The invention is concerned with the systematic elucidation and identification of regulatory sequences. The invention provides among others screenings and detection methods with which regulatory sequences can be identified. The invention further provides regulatory sequences and use thereof in various fields such as, but not limited to protein production, diagnostics, transgenic plants and animals, and the therapeutic field.
Claims77
A method for selecting a DNA sequence with a gene transcription modulating quality, said method comprising:
- providing a transcription system with a variety of fragment-comprising vectors, said vectors further comprising
- i) an element with a gene-transcription repressing quality, and
- ii) a promoter directing transcription of a reporter gene;
- and
- performing a selection step in said transcription system to identify a fragment comprising said DNA sequence with said gene transcription modulating quality.
The method according to any of the preceding claims, wherein the DNA sequence involved in gene-transcription repression is a DNA sequence that is recognized by a protein complex and wherein said transcription system comprises said complex.
The method according to any of the preceding claims, wherein the vector is an episomally replicating vector.
The method according to any of the preceding claims, characterized in that the vector comprises a replication origin from the Epstein-Barr virus (EBV), OriP, and a nuclear antigen (EBNA-1).
A method for identifying a DNA sequence with a gene transcription repressing quality comprising,
- providing a collection of test nucleic acids,
- generating a collection of expression vectors comprising test nucleic acid and a first reporter gene under transcriptional control of a promoter,
- providing cells with said collection of expression vectors,
- selecting a cell or vector containing progeny thereof, wherein transcription of said first reporter gene is repressed, and
- identifying said test nucleic acid in said cell
A method for detecting the presence of a STAR sequence within a nucleic acid sequence of about 50-5000 base pairs, comprising determining the frequency of occurrence in said sequence of at least one sequence pattern and determining that said frequency of occurrence is representative of the frequency of occurrence of said at least one sequence pattern in at least one sequence comprising a STAR sequence.
A method for detecting the presence of a STAR sequence within a nucleic acid sequence of about 50-5000 base pairs, comprising identifying a sequence comprising a STAR sequence in a part of a chromosome of a cell of a species and detecting significant homology between said sequence and a sequence of a chromosome of a different species.
A method for detecting the presence of a STAR element within a nucleic acid sequence of about 50-5000 base pairs of a vertebrate or plant species, comprising identifying whether a flanking sequence of said nucleic acid sequence is conserved in at least one other species.
A library of isolated and/or recombinant nucleic acids comprising a gene transcription modulating quality and/or a gene transcription repressing quality.
A method for selecting a DNA sequence with a gene transcription modulating quality, said method comprising:
- providing a transcription system with a variety of fragment-comprising vectors, said fragment comprising vectors comprising:
- i) an element with a gene transcription modulating quality, and
- ii) a promoter directing transcription of a reporter gene;
- performing a selection step in said transcription system in order to identify a fragment comprising said DNA sequence with said gene transcription modulating quality.
Dependencies are inferred from the claim text.