title: Efficient generation and reversion of chromosomal translocations using CRISPR/Cas technology.
abstract: Chromosomal 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.
  
  
  
  We 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.
  
  
  
  The 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.
authors.affiliation: Horizon Discovery Ltd, Cambridge Research Park, Waterbeach, Cambridge, United Kingdom.
authors.forename: Sergey
authors.initials: S
authors.lastname: Lekomtsev
authors.affiliation: Horizon Genomics GmbH, Campus Vienna Biocenter, Vienna, Austria.
authors.forename: Sofia
authors.initials: S
authors.lastname: Aligianni
authors.affiliation: Horizon Genomics GmbH, Campus Vienna Biocenter, Vienna, Austria.
authors.forename: Ana
authors.initials: A
authors.lastname: Lapao
authors.affiliation: Horizon Genomics GmbH, Campus Vienna Biocenter, Vienna, Austria. t.buerckstuemmer@horizondiscovery.com.
authors.forename: Tilmann
authors.initials: T
authors.lastname: Bürckstümmer
mesh_terms: D064113:CRISPR-Cas Systems; D045744:Cell Line, Tumor; D002471:Cell Transformation, Neoplastic; D016044:Fusion Proteins, bcr-abl / Q000235:genetics; D000072669:Gene Editing; D050939:Gene Fusion; D015321:Gene Rearrangement; D018390:Gene Targeting; D005818:Genetic Engineering; D006801:Humans; D000094704:RNA, Guide, CRISPR-Cas Systems; D014178:Translocation, Genetic
chemical_list: D000094704:RNA, Guide, CRISPR-Cas Systems; D016044:Fusion Proteins, bcr-abl
keywords: BCR-ABL; CD74-ROS1; CRISPR; Cancer; Cas9; Chromosomal rearrangements; Gene fusions; Gene targeting; Philadelphia chromosome; Translocations