Based on this principle, the recently developed DNA-targeting platform, CRISPR-Cas9, has enabled the recruitment of artificial transcription factors (aTFs) to any specific genomic site to induce endogenous gene activation, termed CRISPR activation (CRISPRa) 9, 10, 11, 12, 13. TFs typically consist of two subdomains, a DNA-binding domain (DBD) and an activation domain (AD) which interacts with coactivator complexes to modulate transcriptional level 6, 7, 8. Transcription factors (TFs) ensure this specificity by recognizing and binding to specific DNA sequences to modulate gene expression through their effector domains 4, 5. Regulation of gene expression requires that the transcriptional machinery be precisely and efficiently assembled at specific genomic loci 1, 2, 3. Overall, our work expands the gene activation toolbox for biomedical research. Quantitative imaging illustrated that nascent RNA-directed aTFs could induce the high-density assembly of coactivators at transcription sites, which may explain the larger transcriptional burst size induced by Narta. Moreover, Narta provides better activation potency of some expressed genes than CRISPRa and, when used in combination with CRISPRa, has an enhancing effect on gene activation. Importantly, the activation is reversible, tunable and specific. Using Narta, we demonstrate robust activation of a broad range of exogenous and endogenous genes in various cell types, including zebrafish embryos, mouse and human cells. In contrast to existing methods based on recruiting transcriptional modulators via DNA-binding proteins, we developed a strategy termed Narta ( nascent RNA-guided transcriptional activation) to achieve gene activation by recruiting artificial transcription factors (aTFs) to transcription sites through nascent RNAs of the target gene. Technologies for gene activation are valuable tools for the study of gene functions and have a wide range of potential applications in bioengineering and medicine.
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