CRISPR-mediated targeting of HER2 inhibits cell proliferation through a dominant negative mutation.
With the discovery of the CRISPR/Cas9 technology, genome editing could be performed in a rapid, precise and effective manner. Its potential applications in functional interrogation of cancer-causing genes and cancer therapy have been extensively explored. In this study, we demonstrated the use of the CRISPR/Cas9 system to directly target the oncogene HER2. Directing Cas9 to exons of the HER2 gene inhibited cell growth in breast cancer cell lines that harbor amplification of the HER2 locus. The inhibitory effect was potentiated with the addition of PARP inhibitors. Unexpectedly, CRISPR-induced mutations did not significantly affect the level of HER2 protein expression. Instead, CRISPR targeting appeared to exert its effect through a dominant negative mutation. This HER2 mutant interfered with the MAPK/ERK axis of HER2 downstream signaling. Our work provides a novel mechanism underlying the anti-cancer effects of HER2-targeting by CRISPR/Cas9, which is distinct from the clinical drug Herceptin. In addition, it opens up the possibility that incomplete CRISPR targeting of certain oncogenes could still have therapeutic value by generation of dominant negative mutants.
Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, #04-01 Nanos, Singapore, 138669.
Huajing
H
Wang
Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, #04-01 Nanos, Singapore, 138669. Electronic address: wsun@ibn.a-star.edu.sg.
William
W
Sun
D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D064130:CRISPR-Associated Proteins / Q000235:genetics* / Q000378:metabolism; D064113:CRISPR-Cas Systems; D049109:Cell Proliferation / Q000187:drug effects; D017024:Chemotherapy, Adjuvant; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D005091:Exons; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D018390:Gene Targeting; D015316:Genetic Therapy / Q000379:methods*; D005838:Genotype; D006801:Humans; D061986:MCF-7 Cells; D009154:Mutation; D010641:Phenotype; D000067856:Poly(ADP-ribose) Polymerase Inhibitors / Q000494:pharmacology; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction; D014162:Transfection; D000068878:Trastuzumab / Q000494:pharmacology
D064130:CRISPR-Associated Proteins; D048049:Extracellular Signal-Regulated MAP Kinases; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D018719:Erb-b2 Receptor Tyrosine Kinases; D000068878:Trastuzumab; C508053:ERBB2 protein, human
Breast cancer; Cas9; ERBB2; PARP inhibitor; Therapeutic; Trastuzumab