cDNA

AICSDP-72:TFAM-mEGFP

159745

AICSDP-72:TFAM-mEGFP

Ampicillin

This plasmid has been used with locus-specific CRISPR/Cas9 to add a mEGFP tag to the C-terminus of human TFAM in WTC-11 human induced pluripotent stem cells by the Allen Institute for Cell Science. Linker (AA) sequence: GGGS. After protein tagging using this donor template plasmid and CRISPR/Cas9 reagents, transfected cells may exhibit varying intensity levels of fluorescence, likely due to editing precision. To obtain cells of uniform intensity levels, see our protocol for fluorescence-assisted cell sorting and subcloning of transfected cells (https://www.allencell.org/instructional-videos-and-tutorials-for-cell-methods.html). Further, we recommend PCR-based assays for identifying precisely edited clones as previously described (https://www.molbiolcell.org/doi/abs/10.1091/mbc.e17-03-0209). For more information on the entire plasmid collection, please see https://www.addgene.org/allen-institute-cell-science/ .

Homology arms and linker-mEGFP sequence for C-terminus tagging of human TFAM

37

High Copy