Generation of FOS gene knockout lines from a human embryonic stem cell line using CRISPR/Cas9

DOI: 10.1016/j.scr.2019.101479

Authors: Chunfu Li; Qing Wang; Zhiyong Peng; Yuchen Lin; Huaying Liu; Xiuling Yang; Shan Li; XiaoTing Liu; Jingru Chen

Step 1: iPSC/ESC maintainance

day 0 ~ 1 (duration missing in database; assumed 1 day)

culture “adherent” human ESC in “mTeSR”. “ESC” should be “downregulated” for “FOS”. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table.

Ideally, “ESC” should express “POU5F1”, “SSEA4”, “TRA-1-60”, “TRA-1-81” at the end of culture.

Step 2: endothelial 1

day 1 ~ 9

culture “adherent” human ESC in “unspecified medium” for “8” days. “ESC” should be “downregulated” for “FOS”. Medium should be changed once in “8” days.