CRISPR-based transcription regulators (CRISPR-TRs) have transformed the current synthetic biology landscape by allowing specific activation or repression of any target gene. our strategy relies on the design of stem loop 2 on the single guide RNA (sgRNA). We find individually transcriptional activation of dCas9-VP64 system or dCas9-MCP-p65-HSF1 system is not high, but the combination of the two system vastly improves the effect of transcriptional activation, more than 20 times the ratio of transcriptional activation of dCas9-VP64 system, and green fluorescent protein is used as the output signal, adding an RNA strand for the combination of the two system to respond the small molecule(microRNA, metal ion, mRNA) by strand displacement to visually detect the intracellular environment. Then, we design the complementary sequence of stem loop 2 on the 3’ end or 5’ end of sgRNA, we find the 5’ end of sgRNA tremendously inhibits transcriptional activation, adding the respond sequence between the complementary sequence and sequence of sgRNA to respond a exogenous molecule (DNA oligonucleotide,Csy6A protein)，the ratio of transcriptional activation of responding DNA oligonucleotide or csy6A protein is above 7 or 21 times. Therefore， we offer a modular and versatile framework enabling endogenous and exogenous molecule responding CRISPR-TRs in mammalian cells.

Modular and versatile CRISPR-based transcription regulators responding endogenous and exogenous molecule

Fall 2022 abstracts, posters, and presentations will launch in mid-September.

ACS Fall 2022

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.