Journal: Journal of Cellular and Molecular Medicine

Article Title: Generation of ‘designer erythroblasts’ lacking one or more blood group systems from CRISPR/Cas9 gene‐edited human‐induced pluripotent stem cells

doi: 10.1111/jcmm.16872

Figure Lengend Snippet: CRISPR/Cas9 gene‐edited erythroblasts surface antigen presentation. Complete loss of RhAG, RhD/CE, GPB protein expression was confirmed for RhAG, RhD/CE (A) and GPB (B) erythroblasts. Anti‐U was used to confirm GPB deletion. Kell glycoprotein expression was evaluated as a proxy for XK knockout erythroblasts. Kell expression (C) was considerably diminished consistently with the lack of Kx expression. KO, knockout

Article Snippet: Briefly, cloning was performed by ligating annealed‐guides into BbsI digested GFP‐Cas9 expression vector pX458 (one guide/vector) using a quick ligation kit (New England Biolabs, Ipswich, MA, USA) and transformed into competent E. coli cells (Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: CRISPR, Expressing, Knock-Out