HeLa WT cells
Homo sapiens
Generation of TALEN-mediated LATS2 knockout HeLa-S3 cell line. The ORF of TAL-effector nucleases (TALEN) were synthesized by GeneArt (Life Technologies). Bowtie software was used to confirm whether this target sites are unique in human genome (hg19). The coding region in this entry clone was sub-cloned to expression vector, pDEST26 (Life Technologies) using Gateway technology. Resulted two expression constructs were transfected to HeLa-S3 cells by Lipofectamin 2000 reagent (Life Technologies) followed by manufacturers' instructions. After two days from transfection, transfectant pool was diluted in 96 well plates, then some clones derived from single cell were scaled up and expanded as candidate knockout cell lines. The successful knockout of LATS2 was validated by endonclease assay, sanger sequence of targeted locus and immunoblotting analysis. The most compatible clone with LATS2 knockdown was selected and applied for downstream analyses.
sample_name
HeLa-WT
strain
HeLa-S3
sex
female