Genetic barcodes
View Patent ↗Herein are described multicolor FISH probe sets termed “genetic barcodes” targeting several cancer or disease-related loci to assess gene rearrangements and copy number changes in tumor cells. Two, three or more different fluorophores are used to detect the genetic barcode sections thus permitting unique labeling and multilocus analysis in individual cell nuclei. Gene specific barcodes can be generated and combined to provide both numerical and structural genetic information for these and other pertinent disease associated genes.
1. A method for uniquely labeling a cancer gene of interest in an individual cell nucleus, comprising the steps of:
(a) obtaining and providing a single cell from a patient;
(b) treating the single cell and fixing it to a substrate for analysis to increase accessibility of a target chromosome and to reduce nonspecific binding;
(c) hybridizing a set of three probes to uniquely label the cancer gene of interest in a target chromosome, wherein a first probe is selected to hybridize to the region proximal to the cancer gene on the target chromosome, the second probe is selected to hybridize to the cancer gene of interest, and the third probe is selected to hybridize to the region distal to the cancer gene of interest on the target chromosome,
wherein the set of the three probes provides a unique label for the cancer gene of interest in the individual cell nucleus, and wherein the cancer gene of interest is at least one of Erbbl, Erbb2, HIPK2, PTEN and XPG genes in tumor cells.
2. The method of claim 1 , wherein the three probes are different in size.
3. The method of claim 1 , wherein the three probes are labeled differently from each other.
4. The method of claim 1 wherein the parameters of pattern, size, arrangement and color of the labels provide the unique label of the gene of interest and any detected change in the unique label is an indicator of chromosomal changes such as addition, deletion, amplification, rearrangement and/or translocation.
5. The method of claim 1 , further comprising the step of providing more than one set of chromosome-specific probes to analyze chromosomal changes in genes in tumor cells in human chromosome 4, 7, 10 and 13.
6. The method of claim 5 , wherein the set of chromosome-specific probes comprising a set of three probes each to detect Erbb1, Erbb2, HIPK2, PTEN and XPG genes in tumor cells.
7. A method to detect amplification of a cancer gene, comprising the steps of:
(a) obtaining and providing a single cell from a patient;
(b) treating the single cell and fixing it to a substrate for analysis to increase accessibility of a target chromosome and to reduce nonspecific binding;
(c) providing a set of three probes to detect the amplification of a cancer gene in a target chromosome, wherein a first probe is selected to hybridize to the proximal region of the cancer gene on the target chromosome, the second probe is selected to hybridize to the cancer gene, and the third probe is selected to hybridize to the distal region of the cancer gene on the target chromosome;
(d) hybridizing the probes to the target chromosomes in the single cell;
(e) removing unbound probes; and
(f) detecting the hybridized probes to the target chromosomes, wherein the detection is carried out such that amplification of the cancer gene is detected if hybridization of at least one of probes is detected two or more times.
8. The method of claim 7 , further comprising providing two sets of three probes to detect amplification of two cancer genes.
9. The method of claim 7 , further comprising the steps of:
(g) removing the hybridized probes from the target chromosomes;
(h) repeating steps (a) through (g) for a second and third subset of probes,.
10. The method of claim 7 in which the detecting step is performed using a filter-based fluorescent microscope, optionally equipped with a spectral imaging system.
11. The method of claim 7 in which the detecting step is performed on interphase, non-proliferating or resting cells or organelles.
12. The method of claim 7 in which the detecting step is performed on metaphase cells.