Thymidine: Increase HDR via cell cycle synchronization

CAS #: 50-89-5

Biological Activity

Composed of the pyrimidine base thymine attached to the sugar deoxyribose, thymidine is converted to the corresponding nucleotide prior to in vivo DNA synthesis. In the DNA double helix, thymine base pairs with adenine. As a cell synchronization agent in the laboratory, thymidine is used to (reversibly) arrest cells at the border between the G1 and S phase, preventing DNA synthesis. ^[1]

Successful Applications in CRISPR

(i) Thymidine treatment coupled with timed delivery of pre-assembled Cas9 ribonucleoprotein (RNP) complex, targeting the EMX1 gene in human primary neonatal fibroblasts (neoFB) led to an increase in the rate of total editing (TE) from 5% in unsynchronized cells to 13%. ^[2]

Reference:

1. Chen, G., & Deng, X. (2018). Cell Synchronization by Double Thymidine Block. Bio-protocol, 8(17), e2994. https://doi.org/10.21769/BioProtoc.2994
2. Lin, S., Staahl, B. T., Alla, R. K., & Doudna, J. A. (2014). Enhanced homology-directed human genome engineering by controlled timing of CRISPR/Cas9 delivery. eLife, 3, e04766. https://doi.org/10.7554/eLife.04766

Related Product:

144001 Thymidine, 99% [CAS: 50-89-5]