★ Cas9/gRNA complex binds to proto-spacer adjacent motif(PAM)site and unwinds DNA. ★ gRNA binds to target locus in genomic DNA adjacent to the PAM site. ★ Cas9 cleaves the 5'exons of the gene targeted by the three gRNA plasmids at three specifics sites. ★ Target gene is disrupted.
PARP-1 CRISPR/Cas9 KO Plasmid(h):sc-400046.Western blot analysis of human PARP-1 expression in empty vector control(A) and PARP-1 CRISPR/Cas 9 KO Plasmid transfected(B) Hela whole cell lysates. Blot probed with PARP-1(B-10):sc-74470.α-actinin(H-2):sc-17829 used as specificity and loading control.
HDR(Homology Directed Repair) 修复更精准!
HDR质粒提供用于特异DNA的双链断裂(DSB)修复的模板,比non-homologous end -joining(NHEJ)更精确,不用担心非特异性插入或者缺失裂解位点。
我们的产品:
★ 每个HDR质粒由三条特异性裂解相应CRISPR/Cas9敲除质粒位点的质粒组成 ★ 每个HDR质粒由两条与基因组DNA特异性重组结合800bp的同源arms组成,围绕相应的Cas9引起的双链DNA断裂位点 ★ 每个HDR质粒插入一个嘌呤霉素抗性基因,选择已经发生了Cas9引起的DNA剪切的细胞 ★ 每个HDR质粒包含RFP蛋白,以便确认是否转染 ★ 每个嘌呤毒素抗性基因被两个LoxP位点侧接,以允许由Cre重组载体进行进一步的处理。Provided as transfection-ready,purified plasmid DNA ★ 每个包装20µg,up to 20 transfections
1.Van der Oost J., et al. 2014. Unraveling the Structural and Mechanistic Basis of CRISPR-Cas Systems. Nat. Rev. Microbiol. (7):479-92. PMID 24909109.
7.Sternberger, SH., et al. 2014. DNA Interrogation by the CRISPR RNA-guided Endonuclease Cas9. Nature. 507(7490):62-7. PMID 24476820.
2.Hsu, P., et al. 2014. Development and Applications of CRISPR-Cas9 for Genome Editing. Cell. 157(6):1262-78. PMID 24906146.
8.Gasuinas, G., et al. 2012. Cas9-crRNA Ribonucleoprotein Complex Mediates Specific DNA Cleavage for Adaptive Immunity in Bacteria. Proc. Natl. Acad. Sci. 109(39):E2579-86. PMID 22949671.
3.Nishimasu, H., et al. 2014. Crystal Structure of Cas9 in Complex with Guide RNA and Target DNA. Cell. 156(5):935-49. PMID 24529477.
9.Mali, P., et al. 2013. RNA-Guided Human Genome Engineering via Cas9. Science. 339 (6121): 823-6. PMID 23287722.
4.Deltcheva, E., et al. 2011. CRISPR RNA Maturation by Trans-Encoded Small RNA and Host Factor RNASE III. Nature. 471: 602-607. PMID 21455174.
10.Ran, FA., et al. 2013. Genome engineering using the CRISPR-Cas9 system. Nat. Protoc. (11): 2281-308. doi: 10.1038/nprot.2013.143. PMID 24157548.
5.Jinek, M., et al. 2012. A Programmable Dual-RNA Guided DNA Endonuclease in Adaptive Immunity. Science. 337(6096):816-21. PMID 22745249.
11.Shalem O., et al. 2014. Genome-scale CRISPR-Cas9 Knockout Screening in Human Cells. 343(6166):84-7. PMID 24336571.
6.Deveau, H., et al. 2010. CRISPR/Cas System and its Role in Phage-Bacteria Interaction. Annu. Rev. Microbiol. 64: 475-493. PMID 20528693.
12.Ran, F.A., et al. 2013. Double nicking by RNA-guided CRISPR Cas9 for enhanced genome editing specificity. Cell 154: 1380-1389.