Creation of Philadelphia chromosome by CRISPR/Cas9-mediated double cleavages on BCR and ABL1 genes as a model for initial event in leukemogenesis.
Cancer Gene Ther
; 30(1): 38-50, 2023 01.
Article
in En
| MEDLINE
| ID: mdl-35999358
ABSTRACT
The Philadelphia (Ph) chromosome was the first translocation identified in leukemia. It is supposed to be generated by aberrant ligation between two DNA double-strand breaks (DSBs) at the BCR gene located on chromosome 9q34 and the ABL1 gene located on chromosome 22q11. Thus, mimicking the initiation process of translocation, we induced CRISPR/Cas9-mediated DSBs simultaneously at the breakpoints of the BCR and ABL1 genes in a granulocyte-macrophage colony-stimulating factor (GM-CSF) dependent human leukemia cell line. After transfection of two single guide RNAs (sgRNAs) targeting intron 13 of the BCR gene and intron 1 of the ABL1 gene, a factor-independent subline was obtained. In the subline, p210 BCRABL1 and its reciprocal ABL1BCR fusions were generated as a result of balanced translocation corresponding to the Ph chromosome. Another set of sgRNAs targeting intron 1 of the BCR gene and intron 1 of the ABL1 gene induced a factor-independent subline expressing p190 BCRABL1. Both p210 and p190 BCRABL1 induced factor-independent growth by constitutively activating intracellular signaling pathways for transcriptional regulation of cell cycle progression and cell survival that are usually regulated by GM-CSF. These observations suggested that simultaneous DSBs at the BCR and ABL1 gene breakpoints are initiation events for oncogenesis in Ph+ leukemia. (200/200 words).
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Philadelphia Chromosome
/
Leukemia, Myelogenous, Chronic, BCR-ABL Positive
Type of study:
Prognostic_studies
Limits:
Humans
Language:
En
Journal:
Cancer Gene Ther
Journal subject:
GENETICA MEDICA
/
NEOPLASIAS
/
TERAPEUTICA
Year:
2023
Document type:
Article
Affiliation country: