SCR7
mRNA synthesis
In vitro transcription of capped mRNA with modified nucleotides and Poly(A) tail
Tyramide Signal Amplification (TSA)
TSA (Tyramide Signal Amplification), used for signal amplification of ISH, IHC and IC etc.
Phos Binding Reagent Acrylamide
Separation of phosphorylated and non-phosphorylated proteins without phospho-specific antibody
Cell Counting Kit-8 (CCK-8)
A convenient and sensitive way for cell proliferation assay and cytotoxicity assay
SYBR Safe DNA Gel Stain
Safe and sensitive stain for visualization of DNA or RNA in agarose or acrylamide gels.
Inhibitor Cocktails
Protect the integrity of proteins from multiple proteases and phosphatases for different applications.
Scr7是DNA连接酶IV的抑制剂,最初被认为是一种抗癌剂。
Scr7靶向DNA连接酶IV的DNA结合域,减少其与双链断裂(DSBs)的亲和性并抑制其功能。Scr7也可以抑制DNA连接酶III。用doxycycline处理细胞诱导Cas9的表达,同时用Scr7处理24 h可以使细胞能够进入S/G2期,这对于同源定向修复(HDR)是必须的。用Scr7处理小鼠会影响淋巴细胞的发育,这是由于DNA连接酶IV在V(D)J重组期间通过C-NHEJ16对编码端的接合作用。由于Scr7的非共价结合,淋巴细胞发育的缺陷是暂时并可逆的。在培养的细胞和小鼠中,Scr7通过瞬时阻断NHEJ,从而增加HDR的发生频率,产生精确的基因组编辑。
参考文献:
[1]. Srivastava M, Nambiar M, Sharma S et al. An inhibitor of nonhomologous end-joining abrogates double-strand break repair and impedes cancer progression. Cell. 2012 Dec 21;151(7):1474-87. doi: 10.1016/j.cell.2012.11.054.
[2]. Maruyama T, Dougan SK, Truttmann MC et al.Increasing the efficiency of precise genome editing with CRISPR-Cas9 by inhibition of nonhomologous end joining. Nat Biotechnol. 2015 Mar 23. doi: 10.1038/nbt.3190. [Epub ahead of print]
- 1. Dennis Klug, Katharina Arnold, et al. "A toolbox of engineered mosquito lines to study salivary gland biology and malaria transmission." PLoS Pathog. 2022 Oct 12;18(10):e1010881. PMID: 36223382
- 2. Lampi Y, Van Looveren D, et al. "Targeted editing of the PSIP1 gene encoding LEDGF/p75 protects cells against HIV infection." Sci Rep. 2019 Feb 20; 9 (1): 2389. PMID: 30787394
- 3. Krüger K, Geist K, et al. "Multiple DNA damage-dependent and DNA damage-independent stress responses define the outcome of ATR/Chk1 targeting in medulloblastoma cells." Cancer Lett. 2018 May 16; 430: 34-46. PMID: 29753759
- 4. Fernandez-Godino R, Bujakowska KM, Pierce EA. "Changes in extracellular matrix cause RPE cells to make basal deposits and activate the alternative complement pathway." Hum Mol Genet. 2018 Jan 1; 27 (1): 147-159. PMID: 29095988
- 5. Huberman LB, Coradetti ST, Glass NL. "Network of nutrient-sensing pathways and a conserved kinase cascade integrate osmolarity and carbon sensing in Neurospora crassa." Proc Natl Acad Sci U S A. 2017 Oct 10; 114 (41): E8665-E8674. PMID: 28973881
- 6. Hindriksen S, Bramer AJ, et al. "Baculoviral delivery of CRISPR/Cas9 facilitates efficient genome editing in human cells." PLoS One. 2017 Jun 22; 12 (6): e0179514. PMID: 28640891
- 7. Lee JS, Grav LM, et al. "Accelerated homology-directed targeted integration of transgenes in Chinese hamster ovary cells via CRISPR/Cas9 and fluorescent enrichment." Biotechnol Bioeng. 2016 May 9. PMID: 27159230
Storage | Store at -20°C |
M.Wt | 334.39 |
Cas No. | 1533426-72-0 |
Formula | C18H14N4OS |
Solubility | insoluble in H2O; ≥16.72 mg/mL in DMSO; ≥2.6 mg/mL in EtOH with ultrasonic |
Chemical Name | 5,6-bis((E)-benzylideneamino)-2-thioxo-2,3-dihydropyrimidin-4(1H)-one |
SDF | Download SDF |
Canonical SMILES | S=C(NC(/N=C/C1=CC=CC=C1)=C2/N=C/C3=CC=CC=C3)NC2=O |
运输条件 | 蓝冰运输或根据您的需求运输。 |
一般建议 | 不同厂家不同批次产品溶解度各有差异,仅做参考。若实验所需浓度过大至产品溶解极限,请添加助溶剂助溶或自行调整浓度。溶液形式一般不宜长期储存,请尽快用完。 |
细胞实验: [1] | |
细胞系 |
上皮(A549)和黑色素瘤(MelJuSo)细胞系衍生物 |
制备方法 |
该化合物在DMSO中的溶解度大于10 mM,若配制更高浓度的溶液,一般步骤如下:请将试管置于37℃加热10分钟和/或将其置于超声波浴中震荡一段时间。原液于-20℃可放置数月。 |
反应条件 |
24 小时, 37℃ |
实验结果 |
Scr7增加细胞系中插入诱变的效率。在A549细胞中,相较于未处理的对照组,0.01 μM Scr7将靶位点的插入效率提高约三倍。在Scr7处理的MelJuSo细胞中,插入效率也以剂量依赖性方式增强高达19倍。 |
动物实验: [1] | |
动物模型 |
Kell-LPETG小鼠 |
给药剂量 |
在原核阶段将CRISPR组分混合物(Cas9 mRNA、sgRNA和靶向模板)和10 mM的Scr7 NHEJ抑制剂(至终浓度为1 mM)注射到细胞质中。将注射的受精卵在2-细胞阶段转移到假孕妇中。 |
实验结果 |
共注射Scr7增加了在小鼠胚胎中精确基因组编辑的效率。与未注射Scr7的胚泡相比,使用Scr7共注射的插入效率显著增高(P = 0.0012)。与未注射Scr7的E10胚胎相比,Scr7共注射的E10胚胎中的插入效率也显著增高(P = 0.003)。 |
注意事项 |
请于室内测试所有化合物的溶解度。虽然化合物的实际溶解度可能与其理论值略有不同,但仍处于实验系统误差的允许范围内。 |
References: 1. Maruyama T, Dougan SK, Truttmann MC et al. Increasing the efficiency of precise genome editing with CRISPR-Cas9 by inhibition of nonhomologous end joining. Nat Biotechnol. 2015 May;33(5):538-42. |
质量控制和MSDS
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