Open AccessApplication of Chromosome Conformation Capture Method for Detection MYC/TRD Chromosomal Translocation in Leukemia Cell Line
Author(s) -
Moloud Absalan,
Mohammad Hossein Ghahremani,
Zahra Jabbarpour,
Roya Karimi,
Shilan Shafei,
Reza Heidari,
Mostafa Akbariqomi,
Gholamreza Tavoosidana
Publication year - 2020
Publication title -
international journal of hematology- oncology and stem cell research.
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.436
H-Index - 16
eISSN - 2008-3009
pISSN - 2008-2207
DOI - 10.18502/ijhoscr.v14i3.3729
Subject(s) - chromosomal translocation , biology , breakpoint , microbiology and biotechnology , primer (cosmetics) , chromosome engineering , chromosome , locus (genetics) , gene , genetics , dna , karyotype , chemistry , organic chemistry
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.