*使用本品系发表的文献需注明: R26-CAG-2xKRAB-dCas9-IRES-EGFP mice (Cat. NO. NM-KI-18007) were purchased from Shanghai Model Organisms Center, Inc.. 验证数据 通过Westernblot验证2xKRAB-dCas9表达 Rosa26- CAG-LSL-2xKRAB-dCas9 mice were crossed with Dppa3-Cre mice to delete STOP cassette....
*使用本品系发表的文献需注明: R26-CAG-LSL-dCas9-VPR-IRES-EGFP mice (Cat. NO. NM-KI-18004) were purchased from Shanghai Model Organisms Center, Inc.. 你也可能感兴趣 Tamoxifen诱导Cre-ERT2小鼠 使用指南 Cre-ERT2在无Tamoxifen诱导的情况下,在细胞质内处于无活性状态;当Tamoxifen诱导后,Tamoxifen的代...
To establish a screening platform, we aged cohorts of mice that express Cas9 (and eGFP) in all cells, hereafter termed Cas9 mice49 (Methods). For each independent screen (three in total), we collected NSCs from the SVZ of 6 young (3–4 months old) and 6 old (18–21 months...
因为cas9切割引发egfp读码框移码的概率是相同的,因此,可以根据egfp荧光缺失细胞的比例作为cas9活性的依据。 31.针对egfp grna表达载体构建:针对egfp grna靶位点为:cgatgcccttcagctcgatgcgg(seq id no: 11,粗体下划线部分为pam序列),构建egfp grna表达载体的过程为:通过全基因合成获得包含u6启动子、grna靶序列和grna ...
While BRD0539 is reported to inhibit SpCas9 in the eGFP-disruption assay with an EC50 of 11.5 μM (ref. 16), we failed to observe Cas9 inhibitory effects of BRD0539 and observed cytotoxicity in mESCs at concentrations of 30 μM (Extended Data Fig. 4c). Thus, both anti-CRISPR protein ...
Validation of sgRNA composition and Cas9 protein activity. (a) Endocytosis of EGFP and DiD into HepG2 cells by different nanoparticles for 6 h. Scale bar, 100 nm. (b) Flow cytometry of DiD-OCT/CS/NPs, FITC-BSA-OCT/CS/NPs, and DiD + FITC-BSA-OCT/CS/NPs. (c) Investigation on...
The same group [108] loaded Cas9/sgRNA RNP onto a yarn-like DNA NC, and integrated PEI as an out-layer coating for endo/lysosomal escape for enhanced RNP delivery into U2OS.EGFP (EGFP: enhanced green fluorescent protein) tumor cells. Fig. 3 PEI-based pH-responsive CRISPR-Cas9 delivery ...
For in vitro assessment of gene editing efficiency, the Cas9-mice were crossed with the EGFP-mice to obtain mouse embryonic fibroblasts (MEF) expressing both EGFP and Cas9 (MEF Cas9-EGFP ). Transfection of these cells with in vitro transcribed (IVT) EGFP sgRNA or phU6-EGFP sgRNA plasmid ...
· J South Med Univ, 2019, 39(11): 1381-1386 http://www.j-smu.com Cas9 成为继基因打靶技术,RNAi,ZFNs 和 TALENs 后 又一新兴基因编辑技术[11].张峰团队首次利用改造的 CRISPR/Cas9 系统在人类细胞和小鼠细胞中成功完成 基因组定点编辑[12].南京大学用 CRISPR 系统敲除了小 鼠 EGFP基因,首次将CRISPR...
[67,68]. Notably, the T1337R substitution imparts an additional nucleotide recognition preference, as evidenced by both the EGFP disruption assay and crystal structure analysis, which are in agreement. Furthermore, compared to the wild type, the D1135E substitution enhances specificity by ...