Journal of Stem Cell Research & Therapy

Generation of TUBB3-mCherry reporter knock-in human pluripotent stem cell line (SNUe003-A-4) using CRISPR/Cas9-mediated genome engineering

Joint Event on International Conference on STEM CELL AND ORTHOPEDICS & International Conference on PSYCHOSOMATIC AND LABORATORY MEDICINE

July 19-20, 2023 | Rome, Italy

A-Hyeon Kim

Catholic Kwandong University, Republic of Korea

Posters & Accepted Abstracts: J Stem Cell Res Ther

Abstract:

TUBB3 is a structural neuronal protein important for multiple neuronal functions including axonal guidance and maturation. This study aimed to generate a human pluripotent stem cell (hPSC) line with a TUBB3-mCherry reporter using CRISPR/ SpCas9 nuclease. The stop codon in the last exon of TUBB3 was replaced with a T2A-mCherry cassette using CRISPR/SpCas9- mediated homologous recombination. The established TUBB3-mCherry knock-in cell line exhibited typical pluripotent characteristics. The mCherry reporter faithfully replicated the endogenous level of TUBB3 upon induction of neuronal differentiation. The reporter cell line could contribute to the investigation of neuronal differentiation, neuronal toxicity, and neuronal tracing.

Biography :

A-hyeon Kim completed a master's degree at Catholic Kwandong University in republic of korea. She conducted a study to establish a knock-in cell line using human pluripotent stem cells, and published it in the journal Stem Cell Research. After completing the master's degree, she went to the Ph.D. course at this university and is conducting research on the mechanism of Moyamoya disease, a rare, progressive cerebrovascular disorder caused by blocked arteries at the base of the brain in an area called the basal ganglia. And research on human induced pluripotent stem cells (hiPSCs)-derived red blood cells (RBCs) differentiation is being conducted to express beta-globin and increase mass production and enucleation.