Among diverse genome editing tools, the Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated proteins system (CRISPR/Cas system) has exhibited the obvious advantages of specificity, simplicity, and flexibility over any previous genome editing system. Genome editing technology has become one of the hottest research areas in recent years. (B) SunTag system: dCas9 fused with GCN4 to recruit multiple copies of scFv, TET and other element X to activate the target gene cooperatively (C) dCas9-VPR system: dCas9 fused with VP64-p65-Rta to activate the target gene (D) dCas9-TV system: dCas9 TV activation system includes 6 TAL and 2 VP128 (E) scRNA system: An RNA hairpin domain with RNA sequences MS2, PP7 and com recognized by MCP, PCP and com RNA binding proteins was introduced at the end of sgRNA, and the transcription activating element VP64 was fused into each corresponding RNA binding protein (F) SAM system: The four bases at the distal end of the stem loops of gRNA were modified to recognize the stem loops of MS2 to bind p65 and HSF1 (G) CRISPR-Act2.0. The results suggested that dCas9-6TAL-VP128 was a strong transcriptional activator and was called the dCas9-TV system as shown in Figure 3C. Results showed that the combination of VP128 with a tandem ERF2m-EDLL motif (up to four copies) activated LUC transcription up to 12.6-fold compared to the base level, while combination of VP128 and TALE TAD (up to six copies) increased LUC transcription up to 55-fold.
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