Project titleUsing CRISPR Techniques To Create And Characterize The Phenotype Of Cancer Mutants For HNRPA2B1 And PTPRE Genes

HnRNP A2/B1 gene has many different transcripts, mainly the B1 and A2 transcripts, thanks to alternative splicing, where the difference between the two is the presence or absence of exon 2 which contains the NLS signal. In normal cells, B1 isoform dominates over A2, but recently, it has been shown that the ratios almost flip, making A2 dominate B1 isoform in breast cancer cells, thus making more of the cytosolic version rather than the nuclear one. Little is known about the effect of this major conversion of isoforms. Thus, the first aim of this project is to create cancer mutants that contain A2 or B1 isoforms only to characterize the difference in phenotype between the two. PTPRE shares a similar story, where it's usually mostly membrane bound (memPTPRE isoform), but it has been shown that, in cancer, the cytosolic isoform (cytPTPRE) which lacks the first 3 exons become more prominent, thanks to the existence of a secondary transcription start site (TSS) within the third intron, and little is known about the functions of these isoforms. Therefore, the second aim is to create cancer mutants that contain cytPTPRE or memPTPRE mutants to understand and characterize the difference in phenotype between the two. This project relies on CRISPR techniques, from designing the guide RNAs and selecting the right plasmids (PX458 and PX459) to the Homology-Directed Repair (HDR), to make the necessary modifications to breast cancer cell lines MCF-7 and MDA-MB-231,  to keep or remove exon 2 from the HnRNP A2/B1 gene and disrupt the trascription start sites of PTPRE genes, which will allow the creation of 4 different cancer mutants: B1 only, A2 only, memPTPRE only, and cytPTPRE only mutants. 

Primary contact nameMohammad Hammad
Primary contact emailEmail hidden; Javascript is required.
Primary contact mobile phone+974 7797 3958
Students/participant(s) programs
  • Biological Sciences
Faculty advisor(s)
Advisor name Email Affiliation
Ravichandra Bachu CMU-Q
Ihab Younis CMU-Q
For CMU-Q advisor(s), please select their program(s)
  • Biological Sciences