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Fig.1 Schematic view of CRISPR/Cas9–based editing to create gene knockouts and knockins. [1]
CRISPR/Cas9 technologies have transformed our ability to manipulate genomes for research and gene-based therapy. It is a two-component editing system that includes a Cas9 protein with cleavage activity and a single guide RNA (sgRNA) to bind both Cas9 and target DNA. CRISPR/Cas9 is in principle able to induce double-stranded DNA breaks (DSBs) at any locus 3 base pairs (bp) upstream of an NGG protospacer adjacent motif (PAM). DSBs can be repaired by two major pathways, with the first one being the re-ligation of the broken DNA ends through non-homologous end joining (NHEJ), which is an error-prone pathway that leads to gene knockout by introducing insertions or deletions (indels). The second pathway is homology-directed repair (HDR), in which DSBs are repaired precisely by using homologous DNA sequences as a repair template and creating gene knock-ins. Both single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) can act as efficient HDR templates, however, recent studies demonstrated that single-stranded DNA (ssDNA or ssODN) is the best HDR templates for CRISPR-based gene insertion, replacement, and correction. Compared to dsDNA donors, ssDNAs demonstrated significantly improved editing efficiency and specificity, as well as reduced off-target integration, especially in editing primary cells, and stem cells, and developing transgenic animal models.
Creative Biogene, one of the leading biotechnology companies, can provide high-quality ssDNA preparation kits for customers worldwide to help you produce ssDNA for use as repair templates in knock-in experiments involving CRISPR/Cas9 technology or other genome editing tools.
Reference:
| Cat.No. | Product Name | Price |
|---|---|---|
| SDPK-001 | ssDNA Preparation Kits | Inquiry |
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