通过GFP或者puromycin富集成mixture,然后直接PCR测序或者利用Restriction-fragment length polymorphism (RFLP) 分析方法。 写在最后 目前这中修复基因的方法比较适合在细胞系(包括ES细胞等)/受精卵(Zygote)中操作,因为效率还是比较高的。对于操作细胞系来说,结合单克隆挑取,往往可以获得100%的矫正。对于小鼠受精卵注射操作...
CRISPR protocol-Cas9介导的同源重组 汉恒生物科技(上海)有限公司 400-092-0065 地址：上海市浦东新区蔡伦路150号1号楼2楼 021-*** 邮箱：service@hanbio.net www.hanbio.net Cas9介导的同源重组
Laminar flow downregulates Notch activity to promote lymphatic sprouting. Choi D et al. (2017) J. Clin Investigation. In press. Generation and characterization of tamoxifen-inducible Pax9-CreER Knock-In Mice using CrispR/Cas9. Jifan F et al. (2016) Genesis 54(9):490-496. Efficient delivery ...
6、:JxlQ5 cells/well (50-70% confluency) 500 ng Cas9 (或者 Cas9n ) -sgRNA1M单链同源模板(iorM)三.鉴定方法:通过GFP或者puromycin富集成mixture然后直接PCR测序或者用Restriction-fragmentlength polymoiphism (RFLP)分析方法四 结论:1 WT的Cas9直接切割双链,引起DSB ,重组效果最好,但会51起3錠;2 单链...
In particular, XCas9Y or XYCas9 mice may be crossed to stable sgRNA transgenic lines expressing one or multiple sgRNAs, each transcribed in a ubiquitous, tissue-specific or inducible manner. This greatly facilitates the production of complex multiply gene-deficient animals in order to study ...
61、le-nicking applications (Box 2), along with the 2A- GFP and 2A-Puro fusion constructs (pSpCas9n(BB)-2A-GFP, pSpCas9n(BB)-2A-Puro). In addition to PCR and plasmid-based delivery methods, Cas9 and sgRNAs can be introduced into cells as mRNA and RNA, respectively. Design of repai...
One day later, GFP-positive ES cells were sorted by FACS and cultured in six wells for DNA-FISH analysis. c Representative DNA-FISH analysis of mixed mouse ES cells targeted at either 21-A + B or control sgRNAs. Red, Texas red-labeled whole-chromosome probe for chromosome 21; green...
同时, 该套系统引入了全新的紫外激活GFP检测系统, 可以通过手持式紫外灯直接照射水稻组织, 使后代群 体中的转基因分离检测十分便利.Molecular Plant同 期还发表了中国科学院上海植物逆境生物学研究中 心朱健康研究团队关于单碱基编辑的研究, 该文报 道, 通过融合SpCas9 (D10A)和ecTadA*7.10的方法 实现了对IPA1等...
e.g., cell lines that are hard to transfect, we developedadditional backbone vectors to facilitate selection andscreening for transfected cells. These vectors contain the fl uorescentmaker protein, GFP, or the selectable puromycin resistancegene, linked to the expressing of SaCas9 via a 2A peptide...
After co-transfection, the C2C12 cell lines were sorted by flow cytometry to obtain the cells coexpressing GFP and RFP, and then identified by DNA sequencing. The molecular characteristics of CSE1L were identified by real-time quantitative PCR (RT-qPCR) and Western blo...