Dataset Information


Generation of LAMC2-EGFP knock-in human pancreatic cancer cells

ABSTRACT: Since LAMC2 is a secreted molecule present in the extracellular matrix of the cells, was designed a strategy based on CRISPR/Cas9-mediated homologous recombination to mark LAMC2 cells in human PDACs. A Cas9 single-guide RNAs complementary to sequences overlapping the stop codon of the LAMC2 locus was designed and a donor vector that contained LAMC2 homology arms flanking an EGFP reporter cassette positioned immediately upstream of the stop codon was generated. LF2A self-cleavage peptide in frame with EGFP so that LAMC2-EGFP locus was expressed as a single mRNA was added, whereas the resulting polypeptide was cleaved in the two encoded proteins, LAMC2 and EGFP. L3.6pl and PANC-1 cells were nucleofected with the donor vector together with a guide-RNA-Cas9 (guide). The engineered cells were subcutaneously injected in CD1 male mice and RNA-seq was performed on LAMC2-EGFP+ and EGFP- -derived tumors.

INSTRUMENT(S): NextSeq 500

ORGANISM(S): Homo Sapiens

DISEASE(S): Pancreatic Adenocarcinoma,Pancreatic Adenosquamous Carcinoma

SUBMITTER: Enza Lonardo  

PROVIDER: E-MTAB-11597 | ArrayExpress | 2022-04-11



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