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Användning av RNA-sekvensering för att upptäcka Novel Splice varianter besläktade med drogmotstånd i<em> In Vitro</em> Cancer modeller
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1Department of Pediatric Oncology/Hematology,VU University Medical Center, 2Department of Hematology,VU University Medical Center, 3Department of Medical Oncology,VU University Medical Center, 4Department of Clinical Genetics,VU University Medical Center, 5Division of General and Transplant Surgery, Azienda Ospedaliera Universitaria Pisana,Universita’ di Pisa, 6Amsterdam Immunology and Rheumatology Center,VU University Medical Center, 7Princess Máxima Center for Pediatric Oncology, 8Cancer Pharmacology Lab, AIRC Start-Up Unit,University of Pisa, 9Institute of Nanoscience and Nanotechnology,CNR-Nano
Chapters
- 00:05Title
- 01:09Characterization of Drug Resistance Profiles through Cytotoxicity Assays
- 05:01RNA Isolation and Library Preparation for RNA-sequencing
- 05:55Validation of the Results by RT-PCR and qRT-PCR Assays
- 07:21Results: Detection of Differential Splicing Events in Chemotherapy-resistant Cell Lines Compared to Parental Wild-type by Using RNA-sequencing
- 09:07Conclusion
Här beskriver vi ett protokoll som syftar till att undersöka effekten av avvikande splitsning på läkemedelsresistens i solida tumörer och hematologiska maligniteter. Till detta mål, analyserade vi transcriptomic profiler föräldra och resistenta in vitro-modeller genom RNA-punkter och etablerat en QRT-PCR-baserad metod för att validera kandidatgener.
