Single-stranded DNA (ssDNA) has emerged as a powerful tool in genome engineering, offering researchers precision and flexibility that complement CRISPR technologies. From serving as templates in knock-in experiments to facilitating DNA repair studies, ssDNA is central to advancing both basic research and translational applications.

ssDNA in CRISPR-Mediated Knock-In and Repair

In CRISPR-Cas systems, precise genome modifications often require a donor template for homology-directed repair (HDR). Synthetic ssDNA is widely preferred over double-stranded DNA for several reasons:

● Higher HDR efficiency: The single-stranded structure allows better alignment with the target site.

● Reduced random integration: Minimizing unintended genomic insertions.

● Ease of synthesis: Especially suitable for short to medium-length templates.

Researchers routinely use ssDNA for introducing point mutations, tagging genes with reporters, or repairing pathogenic variants in cellular models. Its versatility makes it an essential reagent for precise genome editing experiments.

Emerging Applications

● Gene therapy research: ssDNA donors enable targeted correction of disease-associated mutations.

● Functional genomics: ssDNA helps study gene function by enabling controlled knock-ins.

● Synthetic biology: ssDNA is used in assembling complex genetic circuits and constructing novel biomolecules.

Technical Considerations

While ssDNA is highly versatile, challenges remain. Longer ssDNA strands are prone to degradation and may require chemical modifications for stability. Efficient delivery into cells or organoids is also critical, and researchers must optimize experimental conditions for maximum editing efficiency.

GenCefe Biotech Solutions

GenCefe Biotech offers custom ssDNA synthesis services, providing high-quality, precisely designed sequences with flexible lengths and modifications. Our solutions support genome editing, HDR studies, and synthetic biology applications, helping researchers achieve reproducible and high-fidelity results in their experiments.