Skip to main content
Egyptian Journal of Medical Human Genetics
Fig. 3 | Egyptian Journal of Medical Human Genetics

Fig. 3

From: B2M gene knockout in HEK293T cells by non-viral delivery of CRISPR-Cas9 system for the generation of universal cells

Fig. 3

A Schematic flowchart of the experimental procedure for knocking out B2M gene out by CRISPR/Cas9 genome editing system. A Generating the desired gRNA through designed guide oligos under temperature condition. B Inserting the gRNA into backbone of PX458 vector (red region) followed by ejection of two-cutting selective sites. C Transforming the resulting vectors into E. coli DH5α and isolating vectors from colonies. D Co-transfecting the collected vectors into HEK293T cell line using commercially transfection reagent, Lipofectamine. E Directing the gRNA into the specific genomic site and editing the targeted DNA through Cas9 protein. F Rejoining the fragment sites by error-prone NHEJ pathway and knocking out B2M gene

Back to article page