ssDNA is not superior to dsDNA as long HDR donors for CRISPR-mediated endogenous gene tagging in human diploid RPE1 and HCT116 cells

• Published in: BMC genomics Vol. 24; no. 1; p. 289
• Main Authors: Mabuchi, Akira, Hata, Shoji, Genova, Mariya, Tei, Chiharu, Ito, Kei K, Hirota, Masayasu, Komori, Takuma, Fukuyama, Masamitsu, Chinen, Takumi, Toyoda, Atsushi, Kitagawa, Daiju
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 29.05.2023; BioMed Central; BMC
• Subjects: Accuracy; Analysis; Cell lines; Cells; Cloning; Clustered Regularly Interspaced Short Palindromic Repeats; CRISPR; CRISPR-Cas; CRISPR-Cas Systems; Diploids; Diploidy; DNA; DNA - metabolism; DNA, Single-Stranded - genetics; Efficiency; Flow cytometry; Fluorescence; Gene Editing - methods; Gene Knock-In Techniques; Genes; Genetic research; Genomes; Genomics; HCT116 Cells; Homology; Humans; Insertion; Integration; Knock-in; Localization; Long ssDNA; Marking; Mutation; Nuclease; Plasmids; Proteins; Repair template; Transgenes; Japan; CRISPR-Cas; Repair template; Long ssDNA; Knock-in

Recent advances in CRISPR technology have enabled us to perform gene knock-in in various species and cell lines. CRISPR-mediated knock-in requires donor DNA which serves as a template for homology-directed repair (HDR). For knock-in of short sequences or base substitutions, ssDNA donors are frequent...