caa a22 4500 606190643 CHVBK 20210128100856.0 cr unu---uuuuu 210128e20151201xx s 000 0 eng 10.1007/s10577-015-9473-9 doi (NATIONALLICENCE)springer-10.1007/s10577-015-9473-9 Multicolor FISHs for simultaneous detection of genes and DNA segments on human chromosomes [Elektronische Daten] [Nobuyoshi Shimizu, Masahiko Maekawa, Satoko Asai, Yoshiko Shimizu] We have developed a convenient multicolor fluorescent in situ hybridization (FISH) (five-, four-, three-, and two-color FISHs) for detecting specific genes/DNA segments on the human chromosomes. As a foundation of multicolor FISH, we first isolated 80 bacterial artificial chromosome (BAC) probes that specifically detect the peri-centromeres (peri-CEN) and subtelomeres (subTEL) of 24 different human chromosomes (nos. 1~22, X, and Y) by screening our homemade BAC library (Keio BAC library) consisting of 200,000 clones. Five-color FISH was performed using human DNA segments specific for peri-CEN or subTEL, which were labeled with five different fluorescent dyes [7-diethylaminocoumarin (DEAC): blue, fluorescein isothiocyanate (FITC): green, rhodamine 6G (R6G): yellow, TexRed: red, and cyanine5 (Cy5): purple]. To observe FISH signals under a fluorescence microscope, five optic filters were carefully chosen to avoid overlapping fluorescence emission. Five-color FISH and four-color FISH enabled us to accurately examine the numerical anomaly of human chromosomes. Three-color FISH using two specific BAC clones, that distinguish 5′ half of oncogene epidermal growth factor receptor (EGFR) from its 3′ half, revealed the amplification and truncation of EGFR in EGFR-overproducing cancer cells. Moreover, two-color FISH readily detected a fusion gene in leukemia cells such as breakpoint cluster region (BCR)/Abelson murine leukemia viral oncogene homologue (ABL) on the Philadelphia (Ph′) chromosome with interchromosomal translocation. Some other successful cases such as trisomy 21 of Down syndrome are presented. Potential applications of multicolor FISH will be discussed. Springer Science+Business Media Dordrecht, 2015 multicolor FISH nationallicence BAC clones nationallicence numerical anomaly nationallicence fusion genes nationallicence trisomy 21 nationallicence ABL : Abelson murine leukemia viral oncogene homologue nationallicence BAC : Bacterial artificial chromosome nationallicence BCR : Breakpoint cluster region, peri-CEN (peri-centromere) nationallicence CML : Chronic myeloid leukemia nationallicence Dyrk1A : Dual specificity tyrosine-phosphorylation-regulated kinase 1A nationallicence EGF : Epidermal growth factor nationallicence EGFR : Epidermal growth factor receptor nationallicence ES : Embryonic stem nationallicence FISH : Fluorescent in situ hybridization nationallicence iPS : Induced pluripotent stem nationallicence PCR : Polymerase chain reaction nationallicence Ph′ : Philadelphia nationallicence TEL : Telomere nationallicence Shimizu Nobuyoshi Keio University, Tsukuba, Ibaraki, Japan aut Maekawa Masahiko Keio University, Tsukuba, Ibaraki, Japan aut Asai Satoko Keio University, Tsukuba, Ibaraki, Japan aut Shimizu Yoshiko Keio University, Tsukuba, Ibaraki, Japan aut Chromosome Research Springer Netherlands 23/4(2015-12-01), 649-662 0967-3849 23:4<649 2015 23 10577 https://doi.org/10.1007/s10577-015-9473-9 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 review-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10577-015-9473-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Shimizu Nobuyoshi Keio University, Tsukuba, Ibaraki, Japan aut NATIONALLICENCE 700 1- Maekawa Masahiko Keio University, Tsukuba, Ibaraki, Japan aut NATIONALLICENCE 700 1- Asai Satoko Keio University, Tsukuba, Ibaraki, Japan aut NATIONALLICENCE 700 1- Shimizu Yoshiko Keio University, Tsukuba, Ibaraki, Japan aut NATIONALLICENCE 773 0- Chromosome Research Springer Netherlands 23/4(2015-12-01), 649-662 0967-3849 23:4<649 2015 23 10577