Cell Gene Editing

Although gene edited animal models are very powerful tools for biomedical research, genetically modified cell lines have many advantages including convenience, fast timeline, and low cost.

In addition to our knockout cell lines, our gene editing services for knockin cell lines include free preliminary evaluation experiments, single cell colonization, genotyping and validation, tumor formation experiments, and FACS analysis.

Application Example #1:

A knockin cell line model was generated with EGFP inserted at the translation initiation site of ACTB (cytoskeleton filiform protein) in the U2OS cell line (human osteosarcoma). 

ACTB gene encodes β-actin, a cytoskeletal protein. The homologous arms of the targeting vector are about 1 kb, and the homologous recombination of the target vector was done by CRISPR/cas9 based EGE™ technology.

A similar gene editing strategy was used for LMNB1, which encodes the nucleoprotein Lamin-B1.

In U2OS cells, flow cytometry plots show that the efficiency of EGFP-ACTB gene knockin mediated by EGE™ (15.02%) is 8 times higher compared to CRISPR/Cas9-mediated knockin (1.91%). In the C6 cell line, the EGE™ system improves the knockin efficiency of EGFP-LMNB1 to 3.6% from 0.19%, which is a 19-fold increase.

Application Example #2:

Gene editing of hESCs and iPSCs

The highly efficient EGE™ system is ideal for editing the genomes of induced pluripotent stem cells (iPSCs) and human embryonic stem cells (hESCs) . 

Multiple iPSC lines are reprogrammed from human fibroblasts by non-integrated plasmids. Panel A shows phase contrast images. This iPSC knockin cell line expresses the pluripotency markers OCT4, SSEA-4 and TRA-1-60 as shown in Panel B and have a normal karyotype (Panel C) . Teratoma forms in immunodeficient mice (Panel D).