RFP negative transgenic cell lines (HEK293T cells containing RFP dead mutant at AAVS1 locus)

CAT#: GE100041C

RFP negative transgenic cell lines (HEK293T cells containing RFP dead mutant at AAVS1 locus), can be edited to convert to RFP positive cells

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Genomic editing of RFP deletion was performed using wild type RFP sequence (oligo donor), and verified gRNA plasmid. The gRNA sequence was designed based on NGG PAM sequence, and cloned in the xCas9 (Cat#GE100078) or SpCas9 (Cat#GE100002) vector. The parent cell used for the gene editing contains a RFP expression cassette which has a big deletion in the coding sequence. 0.5ug of gRNA plasmid and 0.5ug RFP repairing oligo were co-transfected in the RFP negative parent cells. 10 days after transfection, RFP fluorescence can be observed (Left panel) in both xCas9 and SpCas9 transfected cells. However, xCas9 (Top) seems to have a higher editing efficiency than SpCas9 (bottom).

Specifications

Product Data

• Features:
  • RFP negative cells were created by knocking out RFP expression using specific gRNA construct against RFP sequence inserted in the AAVS1 locus of HEK293 cells.
  • The RFP negative cells can be converted into RFP positive cells using validated sgRNA (GE100041G) and rescue donor oligo (GE100041D).