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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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