这项研究中,研究人员首先开发了一种较于以往更加强大的融合dCas9的激活系统(SPH,SunTag-p65-HSF1),并且分别在人和小鼠的细胞里验证了SPH系统的高效性以及特异性(下图)。 在此基础上,研究者构建了受Cre重组酶调控的SPH转基因小鼠,并且证明了在SPH小鼠的原代细胞里导入sgRNA可以激活基因和长链非编码RNA。为了确定该转...
Cre-dependent SunTag-p65-HSF1 (SPH) transgenic mice were generated with piggyBac transposon system in F1 zygotes and crossed with wild type C57BL/6 mice., SPH transgenic mouse containing HA-tagged dCas9 fused with 10xGCN4, which is linked with p65-HSF1 and EGFP in tandem via P2A and ...
We also used the unmodified pCFD3 plasmid for targeting thetwiandengenes in order to compare the effect of dCas9-CBP using gRNAs with or without MS2 loops. The dCas9-VPR activation system (Addgene 78898) was used for comparison, wherein catalytically dead dCas9 (D10A, H39A, H840A, ...
and the promoter-proximal 5′UTR region with an average methylation of 48% as compared to 7% in dCas9 control samples (Fig.1e). In addition to the 416-bp nucleation site (spanned by the 3HER2gRNAs), hypermethylation spread through the downstream CpG island, creating a 1.25-kb hypermethylat...
Despite rapid progresses in the genome-editing field, in vivo simultaneous overexpression of multiple genes remains challenging. We generated a transgenic mouse using an improved dCas9 system that enables simultaneous and precise in vivo transcriptional
Cre-dependent SunTag-p65-HSF1 (SPH) transgenic mice were generated with piggyBac transposon system in F1 zygotes and crossed with wild type C57BL/6 mice., SPH transgenic mouse containing HA-tagged dCas9 fused with 10xGCN4, which is linked with p65-HSF1 and EGFP in tandem via P2A and ...
Cre-dependent SunTag-p65-HSF1 (SPH) transgenic mice were generated with piggyBac transposon system in F1 zygotes and crossed with wild type C57BL/6 mice., SPH transgenic mouse containing HA-tagged dCas9 fused with 10xGCN4, which is linked with p65-HSF1 a
After fusing with activators, such as p65 and HSF1 transactivation domains, this SAM complex will increase the recruitment of transcription factors and subsequently activate the endogenous gene expression up to 105-fold [119]. When the SAM strategy was introduced into Arabidopsis plants, Park et al...
Judging from the fact that molecular weight of TET1 is greater than all the factor X (VP64, p65HSF1, p300, SS18, GADD45A, FOXA1, and PU.1) including VP64 which is the smallest among the factor X, 22-amino acid linker would also give the best result in the SunTag system with ...