Endogenous N-terminal 3x FLAG Tagging of SLC45A1 in iPSCs

This protocol describes the steps for endogenously tagging the N-terminal of the SLC45A1 gene in WTC11-Ngn2 iPSCs with a 3X FLAG tag. It utilizes a Cas9-sgRNA ribonucleoprotein (RNP) complex and a single-stranded donor oligonucleotide (ssODN) template for homology-directed repair (HDR). The procedure can be adapted for generating point mutations or inserting sequences up to 130 bp. In this case, a 66 bp 3X FLAG tag is inserted after the start codon