Interrogating Cancer Drivers via CRISPR – Webinar
April 1, 2023 - February 23, 2024
Interrogating Cancer Drivers via CRISPR: Mutant TP53 as a Therapeutic Target in Hypodiploid B-cell Acute Lymphoblastic Leukemia
You are invited to a free webinar featuring Dr. Ernesto Diaz-Flores, a pediatric oncologist from the University of California at San Francisco. In this one hour telecast, Dr. Diaz-Flores will speak to the challenges and opportunities behind creating isogenic, hard-to-transduce hypodiploid cell lines using CRISPR technology as well as how to transduce primary cells. You will learn how to ensure high editing efficiency as well as how early selection of purified viable cells can greatly enhance downstream applications, including the generation of patient-derived xenografts.
In the last few years, interrogation of the role of altered genes in cancer has been greatly facilitated by the use of CRISPR/Cas. This gene-editing approach is a key research tool for modulating gene expression (knockout and knockin) efficiently and precisely. However, this approach can be limited depending on the cell type (hard-to-transduce, or primary cells). Such limitations are a challenging roadblock to advancing studies of a wide spectrum of malignant cells.
Patients with hypodiploid B-cell acute lymphoblastic leukemia (B-ALL) present with very poor prognosis (5-year event free survival <29%). These cells have been shown to exhibit mutations in p53 in >90% of cases (Diaz-Flores et al., Cancer Research, 2019). To better understand the mechanism of resistance that arises from p53 mutations, we wished to generate isogenic hypodiploid cell lines (including NALM-16) deficient in p53 and with knocking alleles of p53 wildtype or different mutant versions.
This seminar will cover the steps taken to go from 1%–2% lentiviral transduction efficiency to >95% in hard-to-transduce hypodiploid cell lines and primary cells. We will present how we were able to quickly and gently enrich transduced clones to allow such high transduction efficiency and the generation of knockouts for further studies.