Generation of KRAS knockout pancreatic cancer cell line PANC1
收藏DataONE2024-01-17 更新2024-06-08 收录
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We used CRISPR to inactivate mutant KRAS and STAT3 in PANC1 (KRASG12D) pancreatic cancer cell line. Gene expression analysis of KRAS intact vs. knockout cells identified sets of genes involved in protein synthesis, cell differentiation, and metabolic processes, while the expression of MAPK/ERK target genes remained unperturbed., , , # PANC1 pancreatic cancer cell line
We used CRISPR to inactivate mutant KRAS and STAT3 in PANC1 (KRASG12D) pancreatic cancer cell line. Gene expression analysis of KRAS intact vs. knockout cells identified sets of genes involved in protein synthesis, cell differentiation, and metabolic processes, while the expression of MAPK/ERK target genes remained unperturbed.
Methods
Cell line construction
For CRISPR/Cas9-mediated knockouts, we used sgRNAs specific for KRAS and STAT3. Knockout efficiency was measured by Western blotting. Multiple technical replicates were obtained for each CRISPR targeted gene. Genomic DNA from clonal cell lines was collected using QiaAmp DNA Mini Kit (Qiagen). PCR products for sequencing were amplified using Taq DNA polymerase (Invitrogen) and cloned into a pBluescript vector. At least 10-15 bacterial colonies were sequenced per cell line. For RNA isolation, cells were harvested with TRIzol reagent (Invitrogen). Cells were maintained in DME medium supplemente...
创建时间：
2025-07-26
