Development of a Fluorescence in Situ Hybridization Probe for Detecting IKZF1 Deletion Mutations in Patients with Acute Lymphoblastic Leukemia
| Autor: | Kaoru Kahata, Satoshi Oguri, Masao Nakagawa, Chikara Shimizu, Takanori Teshima, Junichi Hashiguchi, Takeshi Kondo, Masahisa Tsuji, Masahiro Onozawa, Daigo Hashimoto, Kohei Okada, Shinichi Fujisawa |
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| Rok vydání: | 2018 |
| Předmět: |
RAG1/2-mediated rearrangement
Adult Male clone (Java method) Biology Pathology and Forensic Medicine Ikaros Transcription Factor Young Adult 03 medical and health sciences Exon 0302 clinical medicine FISH ikaros Cell Line Tumor medicine Humans Recombination signal sequences Child Interphase Gene In Situ Hybridization Fluorescence Aged Sequence Deletion Aged 80 and over Bacterial artificial chromosome Base Sequence medicine.diagnostic_test illegitimate V(D)J rearrangement Inverse polymerase chain reaction Chromosome RNA Probes Middle Aged Precursor Cell Lymphoblastic Leukemia-Lymphoma IKZF1 Molecular biology 030220 oncology & carcinogenesis Molecular Medicine Female 030215 immunology Fluorescence in situ hybridization |
| Zdroj: | The Journal of Molecular Diagnostics. 20:446-454 |
| ISSN: | 1525-1578 |
| Popis: | Intragenic deletion of IKZF1 is a recurrent genomic alteration in acute lymphoblastic leukemia. The deletions are mediated by illegitimate variable(diversity)joining recombination via cryptic recombination signal sequences (RSSs). We developed a fluorescence in situ hybridization (FISH) probe set that can detect any type of IKZF1 deletion, including the commonly deleted exon 4 to 7 region. The probe set consists of a designed probe for the commonly deleted region (Cy3; red) and a bacterial artificial chromosomes clone probe for detecting the 3' flanking region (Spectrum Green). Intact IKZF1 showed a fusion signal, and the deleted allele showed loss of the red signal (0R1G1F). The FISH probes worked correctly for human leukemic cell lines and clinical samples. One case showed an atypical break-apart signal (1R1G1F). Inverse PCR of the case revealed rearrangement of the excised IKZF1 fragment into a legitimate RSS site at Ig κ on chromosome 2, suggesting a pathogenic role of this recombination-activating gene 1/2-mediated event. In this study, we established FISH probe detecting IKZF1 deletion in a quick, quantitative, and cost-effective manner, and the results provided a novel insight into B-cell receptor editing by rearrangement of a cryptic RSS-mediated genomic fragment in acute lymphoblastic leukemia pathology. |
| Databáze: | OpenAIRE |
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