Recent research highlights the effectiveness of single-stranded DNA (ssDNA) as a homology-directed repair donor template in CRISPR gene editing. Compared to double-stranded DNA (dsDNA), ssDNA significantly boosts editing efficiency, reduces off-target effects, and is versatile in various biological reactions, especially in DNA nanotechnology.
In CRISPR and CRISPR-Cas9 genome editing experiments, long ssDNA serves as a potent donor template, enhancing both insertion and gene replacement efficiency. Its applications extend to single-strand conformation polymorphism, in vitro transcription studies, nucleic acid enzyme S1 mapping, probe preparation, labeling, and differential hybridization.
Beyond experiments, ssDNA plays a role in DNA nanotechnology, serving as a scaffold for drug delivery, molecular diagnostics, DNA-based data storage, and diverse nanoscale applications. With a lower risk of random integration, ssDNA is particularly suitable for gene editing in primary cells, stem cells, and the creation of genetically modified animal models.
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