Application of Chromosome Conformation Capture Method for Detection Myc/Trd Chromosomal Translocation in Leukemia Cell Line Publisher



Absalan M¹ ; Ghahremani MH² ; Jabbarpour Z¹ ; Karimi R³ ; Shafei S¹ ; Heidari R¹ ; Akbariqomi M¹ ; Tavoosidana G¹ Show Affiliations
Source: International Journal of Hematology-Oncology and Stem Cell Research Published:2020

Abstract

Background: Chromosomal breakpoints are the most common cause of hereditary diseases and cancers. Today, many standard clinical methods such as cytogenetic and PCR based techniques are used which have limitation regarding detection resolution. Chromosome conformation capture is a method for detecting gene proximity and chromosomal rearrangements. Materials and Methods: In this study, SKW3 cell line was used for detecting t(8;14)(q24;q11) using a 3C-based technique. SKW3 cell line was used for 3C library preparation. For Inverse PCR, two regions were selected in upstream and downstream of the viewpoint locus on chromosome 8-MYC gene based on EcoRI restriction sites. The captured sequence with intra-chromosomal interaction between chr8-c-MYC and chr14-TRD was selected for the translocation PCR primer design. Results: The DNA fragment captured in 3C PCR showed a specific TRD sequence translocated downstream of the MYC gene. Translocation PCR demonstrated the existence of (8; 14) (q24; q11) MYC /TRD in both library and genomic DNA. Conclusion: This result demonstrated 3C-based method could be used as a useful low-cost easy operating technique in chromosomal rearrangements detection. In this study, the integration of whole genome library monitoring and PCR method was used as a high-through put method in chromosomal breakpoints detection. © 2020, Tehran University of Medical Sciences (TUMS). All rights reserved.