SAMD00021088
External Id
Kyushu University School of Medicine SSP Stem Cell Unit.
INSDC center name
2016-08-10T23:18:47Z
INSDC first public
2016-08-10T10:18:47Z
INSDC last update
DRS033314
INSDC secondary accession
live
INSDC status
Generic
NCBI submission model
Generic.1.0
NCBI submission package
DRS033314
SRA accession
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.
description
Homo sapiens
http://purl.obolibrary.org/obo/NCBITaxon_9606
organism
HeLa-WT
sample name
female
http://purl.obolibrary.org/obo/PATO_0000383
sex
HeLa-S3
http://www.ebi.ac.uk/efo/EFO_0002791
strain
HeLa WT cells
title