接下来利用crispr的载体质粒px458上带有gfp荧光蛋白的特点对这些细胞进行进一步的富集。将共转染crispr和donor后培养24小时的细胞进行流式分选,筛选出gfp阳性的细胞,将这些细胞继续培养,待长满后再次用流式细胞仪分选出dsred阳性的细胞。提取基因组并分别对两侧边界序列及中间的插入片断进行巢式pcr扩增,琼脂糖凝胶电泳检...
GFP的发现引发了生物学领域的“绿色革命”,其基因序列被成功解析,并在大肠杆菌、秀丽线虫和小鼠等非原生生物中实现了表达[1]。科学家们随后发现了YFP、RFP、EGFP、DsRed、mCherry和tdTomato等荧光蛋白的变种或新型荧光蛋白,并通过将这些荧光蛋白与目标基因融合,成功地在大小鼠体内实现了基因转录的跟踪和蛋白质的定位,...
GFP的发现引发了生物学领域的“绿色革命”,其基因序列被成功解析,并在大肠杆菌、秀丽线虫和小鼠等非原生生物中实现了表达[1]。科学家们随后发现了YFP、RFP、EGFP、DsRed、mCherry和tdTomato等荧光蛋白的变种或新型荧光蛋白,并通过将这些荧光蛋白与目标基因融合,成功地在大小鼠体内实现了基因转录的跟踪和蛋白质的定位,...
GzmbCre;Rosa26ACTB-tdTomato, -EGFP/+ mice have a low frequency of GFP+ mature NK cells.Xu, YiyingEvaristo, CésarAlegre, MariaLuisaGurbuxani, SandeepL. Kee, Barbara
Because the administration of a high dose of tamoxifen to Lgr5-DTR-GFP mice led to the expression of Lgr5 and Gpr30 in the upper mucosal region and thus raised the question of selective chief cell labeling, the authors replaced the tamoxifen-dependent Cre/LoxP system with a tamoxifen-...
PCR analysis confirmed the correct removal of the Neo gene and the inversion of iEGFP in the pRosa26-EGFP cells (Supplementary information, Figure S4A). Karyotype analysis showed that these cells contained normal chromosomes (Supple- mentary information, Figure S4B). The other surrogate mother was...
3) ROSA26 TALENs (or MCP-ROSA26-CG01) + donor in vector DC-DON-SH02 Select cells in Puromycin for 7-10 days, after which colonies should be GFP positive. Expression of the insert may be detected by qPCR or Western blot. 4) Donor in vector DC-DON-SH02 only Select cells in ...
Imayoshi, I., Tabuchi, S., Matsumoto, M.et al.Light-induced silencing of neural activity in Rosa26 knock-in and BAC transgenic mice conditionally expressing the microbial halorhodopsin eNpHR3.Sci Rep10, 3191 (2020). https://doi.org/10.1038/s41598-020-59984-3 ...
Genetically modified mousemodels are indispensable tools to reveal principal mechanisms of gene functions. Homologous recombination (HR) inES cellsis routinely used to manipulate genes in mice[1]. However, establishment of anES cellsline with a disrupted or modified “gene of interest” includes several...
GFP的发现引发了生物学领域的“绿色革命”,其基因序列被成功解析,并在大肠杆菌、秀丽线虫和小鼠等非原生生物中实现了表达[1]。科学家们随后发现了YFP、RFP、EGFP、DsRed、mCherry和tdTomato等荧光蛋白的变种或新型荧光蛋白,并通过将这些荧光蛋白与目标基因融合,成功地在大小鼠体内实现了基因转录的跟踪和蛋白质的定位...