Efficient generation and reversion of chromosomal translocations using CRISPR/Cas technology

BMC Genomics

Research Article

Sergey Lekomtsev¹  Tilmann Bürckstümmer²  Sofia Aligianni²  Ana Lapao² 

[1]Horizon Discovery Ltd, Cambridge Research Park, Waterbeach, Cambridge, United Kingdom

[2]Horizon Genomics GmbH, Campus Vienna Biocenter, Vienna, Austria

关键词: Gene fusions;    Translocations;    Chromosomal rearrangements;    CRISPR;    Cas9;    Gene targeting;    Philadelphia chromosome;    BCR-ABL;    CD74-ROS1;    Cancer;   

DOI  :  10.1186/s12864-016-3084-5

 received in 2015-12-27, accepted in 2016-09-14,  发布年份 2016

来源: Springer

【 摘 要 】

BackgroundChromosomal translocations are a hallmark of cancer cells and give rise to fusion oncogenes. To gain insight into the mechanisms governing tumorigenesis, adequate model cell lines are required.ResultsWe employ the versatile CRISPR/Cas system to engineer cell lines in which chromosomal translocations are either generated de novo (CD74-ROS1) or existing translocations are reverted back to the original configuration (BCR-ABL1). To this end, we co-apply two guide RNAs to artificially generate two breakpoints and screen for spontaneous fusion events by PCR.ConclusionsThe approach we use is efficient and delivers clones bearing translocationsin a predictable fashion. Detailed analysis suggests that the clones display no additional undesired alterations, implying that the approach is robust and precise.

【 授权许可】

CC BY   
© The Author(s). 2016