RRC ID 65447
Author Tsukamoto S, Nakade K, Wakabayashi T, Nakashima K, Takami M, Hemmi Y, Kuramochi Y, Shimizu T, Arai Y, Matsuo-Takasaki M, Noguchi M, Nakamura Y, Miwa Y, Hayashi Y.
Title Generation of two ISL1-tdTomato reporter human induced pluripotent stem cell lines using CRISPR-Cas9 genome editing.
Journal Stem Cell Res
Abstract ISL1 encodes a member of the LIM/homeodomain family of transcription factors. This encoded protein plays central roles in the development of motor neuron, pancreas, and secondary heart field. Here we generated heterozygous fluorescent reporters of the ISL1 gene in human induced pluripotent stem cells (hiPSCs). CRISPR/Cas9 genome editing technology was employed to knock-in 2A-tdTomato and EF1 alpha promoter-driven Bleomycin resistance gene to the translational ISL1 C-terminal region. The resulting ISL1-TEZ lines showed tdTomato fluorescence upon motor neuron differentiation. These reporter iPSC lines provide opportunity for monitoring and purifying these related cell lineages.
Volume 53
Pages 102363
Published 2021-5-1
DOI 10.1016/j.scr.2021.102363
PII S1873-5061(21)00209-9
PMID 34087992
MeSH CRISPR-Cas Systems / genetics Clustered Regularly Interspaced Short Palindromic Repeats Gene Editing* Humans Induced Pluripotent Stem Cells* Luminescent Proteins
IF 4.495
Resource
Human and Animal Cells 1383D6(HPS1006) Vero(RCB0001)
DNA material pUC GEB-EcoRV (RDB17460) pUC TEBL-EcoRV (RDB17461) pUC GEB (RDB17462) pUC tk2 (RDB17468) pCRTK2x2-ISL1TEBl (RDB18669) pCRtk2x2NN (RDB18670) pX330 gIsl1 (RDB18671) pUC-TEZ (RDB18672)