scholarly journals Highly-efficient Cas9-mediated transcriptional programming

2014 ◽  
Author(s):  
Alejandro Chavez ◽  
Jonathan Scheiman ◽  
Suhani Vora ◽  
Benjamin W Pruitt ◽  
Marcelle Tuttle ◽  
...  
Keyword(s):  
Transcriptional Activation ◽  
Pluripotent Stem Cells ◽  
Rational Design ◽  
Gene Activation ◽  
Transcriptional Regulator ◽  
Activation Domain ◽  
Transcriptional Activation Domain ◽  
Cas9 Protein ◽  
Potential Applications ◽  
Induced Pluripotent

The RNA-guided bacterial nuclease Cas9 can be reengineered as a programmable transcription factor by a series of changes to the Cas9 protein in addition to the fusion of a transcriptional activation domain (AD). However, the modest levels of gene activation achieved by current Cas9 activators have limited their potential applications. Here we describe the development of an improved transcriptional regulator through the rational design of a tripartite activator, VP64-p65-Rta (VPR), fused to Cas9. We demonstrate its utility in activating expression of endogenous coding and non-coding genes, targeting several genes simultaneously and stimulating neuronal differentiation of induced pluripotent stem cells (iPSCs).

scholarly journals Gene Activation by Dissociation of an Inhibitor from a Transcriptional Activation Domain

2009 ◽  
Vol 29 (20) ◽  
pp. 5604-5610 ◽  
Author(s):  
Fenglei Jiang ◽  
Benjamin R. Frey ◽  
Margery L. Evans ◽  
Jordan C. Friel ◽  
James E. Hopper
Keyword(s):  
Gene Expression ◽  
Transcriptional Activation ◽  
Fluorescent Protein ◽  
Gene Activation ◽  
Gene Array ◽  
Strong Support ◽  
Live Cells ◽  
Activation Domain ◽  
Transcriptional Activation Domain ◽  
Gene Switch

ABSTRACT Gal4 is a prototypical eukaryotic transcriptional activator whose recruitment function is inhibited in the absence of galactose by the Gal80 protein through masking of its transcriptional activation domain (AD). A long-standing nondissociation model posits that galactose-activated Gal3 interacts with Gal4-bound Gal80 at the promoter, yielding a tripartite Gal3-Gal80-Gal4 complex with altered Gal80-Gal4 conformation to enable Gal4 AD activity. Some recent data challenge this model, whereas other recent data support the model. To address this controversy, we imaged fluorescent-protein-tagged Gal80, Gal4, and Gal3 in live cells containing a novel GAL gene array. We find that Gal80 rapidly dissociates from Gal4 in response to galactose. Importantly, this dissociation is Gal3 dependent and concurrent with Gal4-activated GAL gene expression. When galactose-triggered dissociation is followed by galactose depletion, preexisting Gal80 reassociates with Gal4, indicating that sequestration of Gal80 by Gal3 contributes to the observed Gal80-Gal4 dissociation. Moreover, the ratio of nuclear Gal80 to cytoplasmic Gal80 decreases in response to Gal80-Gal3 interaction. Taken together, these and other results provide strong support for a GAL gene switch model wherein Gal80 rapidly dissociates from Gal4 through a mechanism that involves sequestration of Gal80 by galactose-activated Gal3.

scholarly journals Identification of a novel efficient transcriptional activation domain from Chinese fir (Cunninghamia lanceolata)

2020 ◽  
Author(s):  
Tengfei Zhu ◽  
Wenyu Tang ◽  
Delan Chen ◽  
Renhua Zheng ◽  
Jian Li ◽  
...  
Keyword(s):  
Transcriptional Activation ◽  
Transcriptional Repression ◽  
High Efficiency ◽  
Gene Activation ◽  
Chinese Fir ◽  
Luciferase Assay ◽  
Virus Protein ◽  
Activation Domain ◽  
Transcriptional Activation Domain ◽  
Flowering Locus

AbstractActivation domains are used as critical components of artificial gene modification tools for genetic breeding. The high efficiency of the activation domain relies on the host plant. However, no activation domain has been identified that originates from Chinese fir (Cunninghamia lanceolate). In this study, a novel strong activator was identified from the whole Chinese fir cDNA library. This plant conserved activator was named TAC 3 (Transcriptional Activation domain from Chinese fir 3). C-terminal 70 amino acids of TAC (TAC3d) have a stronger ability than the commonly used strong activation domain of the virus protein VP16, or the strong plant activation domain, EDLL, in Chinese fir. Through Dual-luciferase assay, phenomic analysis and FT (Flowering Locus T [FT]) quantification, it was shown that, TAC3d can overcome the transcriptional repression of strong plant repressors (Flowering Locus C [FLC]) when fused to its C-terminal domain, thus inhibit the repression of FT expression. In conclusion, for the first time, an activation domain has been identified from Chinese fir. TAC3, which can be used for precise gene activation in Chinese fir in the future, and its function in the plant is more powerful than the commonly used strong activation domain (such as VP16 and EDLL).HighlightTAC3 is the first transcriptional activation domain identified from Chinese fir and its function is more powerful than some commonly used strong transcriptional activators (such as VP16 and EDLL)

scholarly journals Fos and jun cooperate in transcriptional regulation via heterologous activation domains.

1990 ◽  
Vol 10 (10) ◽  
pp. 5532-5535 ◽  
Author(s):  
C Abate ◽  
D Luk ◽  
E Gagne ◽  
R G Roeder ◽  
T Curran
Keyword(s):  
Gene Expression ◽  
Transcriptional Activation ◽  
Transcriptional Activity ◽  
Regulation Of Gene Expression ◽  
Negative Influence ◽  
Activation Domain ◽  
Activation Domains ◽  
Transcriptional Activation Domain ◽  
Transcriptional Stimulation

The products of c-fos and c-jun (Fos and Jun) function in gene regulation by interacting with the AP-1 binding site. Here we have examined the contribution of Fos and Jun toward transcriptional activity by using Fos and Jun polypeptides purified from Escherichia coli. Fos contained a transcriptional activation domain as well as a region which exerted a negative influence on transcriptional activity in vitro. Moreover, distinct activation domains in both Fos and Jun functioned cooperatively in transcriptional stimulation. Thus, regulation of gene expression by Fos and Jun results from an integration of several functional domains in a bimolecular complex.

Transcriptional Activation Domain of Human BTEB2, A GC Box-Binding Factor

1997 ◽  
Vol 121 (2) ◽  
pp. 389-396 ◽  
Author(s):  
S. Kojima ◽  
A. Kobayashi ◽  
O. Gotoh ◽  
Y. Ohkuma ◽  
Y. Fujii-Kuriyama ◽  
...  
Keyword(s):  
Transcriptional Activation ◽  
Activation Domain ◽  
Transcriptional Activation Domain ◽  
Binding Factor ◽  
Gc Box

Frazzled/Dcc acts independently of Netrin to promote germline survival during Drosophila oogenesis

Development ◽  
2021 ◽  
Vol 148 (24) ◽  
Author(s):  
Samantha A. Russell ◽  
Kaitlin M. Laws ◽  
Greg J. Bashaw
Keyword(s):  
Cell Death ◽  
Axon Guidance ◽  
Transcriptional Activation ◽  
Nutrient Sensing ◽  
Activation Domain ◽  
Transcriptional Activation Domain ◽  
Egg Chambers ◽  
Death Effector ◽  
Female Germline

ABSTRACT The Netrin receptor Frazzled/Dcc (Fra in Drosophila) functions in diverse tissue contexts to regulate cell migration, axon guidance and cell survival. Fra signals in response to Netrin to regulate the cytoskeleton and also acts independently of Netrin to directly regulate transcription during axon guidance in Drosophila. In other contexts, Dcc acts as a tumor suppressor by directly promoting apoptosis. In this study, we report that Fra is required in the Drosophila female germline for the progression of egg chambers through mid-oogenesis. Loss of Fra in the germline, but not the somatic cells of the ovary, results in the degeneration of egg chambers. Although a failure in nutrient sensing and disruptions in egg chamber polarity can result in degeneration at mid-oogenesis, these factors do not appear to be affected in fra germline mutants. However, similar to the degeneration that occurs in those contexts, the cell death effector Dcp-1 is activated in fra germline mutants. The function of Fra in the female germline is independent of Netrin and requires the transcriptional activation domain of Fra. In contrast to the role of Dcc in promoting cell death, our observations reveal a role for Fra in regulating germline survival by inhibiting apoptosis.

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