Source 1primary paper2016Frontiers in Molecular Neuroscience
Toolkit/H1/TO promoter
H1/TO promoter
Also known as: H1/TO promoters
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The H1/TO promoter is a tetracycline-operator-modified H1 RNA polymerase III promoter used in a multi-component CRISPR/Cas9 switch to drive doxycycline-dependent guide RNA expression. In the reported system, it functioned with TetR-mediated regulation and supported in vitro DNA editing with efficiency similar to a U6/TO promoter.
Usefulness & Problems
Why this is useful
This promoter is useful for building doxycycline-responsive CRISPR/Cas9 systems in which gRNA expression can be externally controlled. The cited study positions it as a component for inducible genome editing and suggests potential compatibility with existing Streptococcus pyogenes Cas9 systems, although that compatibility was presented as a possibility rather than a demonstrated result.
Source:
We also demonstrate that our inducible gRNAi vector can be used to edit the genomes of neurons in vivo within the mouse brain in a Dox dependent manner.
Source:
Here, we report the development of a viral mediated CRISPR/Cas9 system that can be rendered inducible utilizing doxycycline (Dox) and can be delivered to cells in vitro and in vivo utilizing adeno-associated virus (AAV).
Problem solved
It addresses the problem of making gRNA expression, and therefore CRISPR/Cas9 editing activity, conditional on doxycycline rather than constitutive. This is specifically relevant for multi-component viral genome-editing systems that require regulated RNA polymerase III-driven gRNA transcription.
Source:
Here, we report the development of a viral mediated CRISPR/Cas9 system that can be rendered inducible utilizing doxycycline (Dox) and can be delivered to cells in vitro and in vivo utilizing adeno-associated virus (AAV).
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Mechanisms
crispr/cas9 genome editingcrispr/cas9-mediated dna editingsmall-molecule-inducible transcriptional derepressionsmall-molecule-inducible transcriptional repression/derepressiontetracycline-operator regulationtetr-operator regulationTechniques
Computational DesignTarget processes
No target processes tagged yet.
Implementation Constraints
The reported inducible gRNA vector was designed to express the gRNA from an H1/TO promoter and TetR for doxycycline-dependent regulation. The promoter is therefore implemented as part of a multi-component system requiring TetR and doxycycline, and it was used in the context of AAV vector design; the supplied evidence does not specify exact construct architecture beyond these elements.
The supplied evidence is limited to one 2016 study and one explicit performance statement, so breadth of validation is narrow. The available text does not provide quantitative editing rates, promoter sequence details, leakiness measurements, kinetics, or independent replication, and the claimed broader compatibility with existing S. pyogenes Cas9 systems remains hypothetical in the provided evidence.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
H1/TO promoters of varying length and a U6/TO promoter can edit DNA with similar efficiency in vitro in a doxycycline-dependent manner.
We show that H1/TO promoters of varying length and a U6/TO promoter can edit DNA with similar efficiency in vitro, in a Dox dependent manner.
H1/TO promoters of varying length and a U6/TO promoter can edit DNA with similar efficiency in vitro in a doxycycline-dependent manner.
We show that H1/TO promoters of varying length and a U6/TO promoter can edit DNA with similar efficiency in vitro, in a Dox dependent manner.
H1/TO promoters of varying length and a U6/TO promoter can edit DNA with similar efficiency in vitro in a doxycycline-dependent manner.
We show that H1/TO promoters of varying length and a U6/TO promoter can edit DNA with similar efficiency in vitro, in a Dox dependent manner.
H1/TO promoters of varying length and a U6/TO promoter can edit DNA with similar efficiency in vitro in a doxycycline-dependent manner.
We show that H1/TO promoters of varying length and a U6/TO promoter can edit DNA with similar efficiency in vitro, in a Dox dependent manner.
H1/TO promoters of varying length and a U6/TO promoter can edit DNA with similar efficiency in vitro in a doxycycline-dependent manner.
We show that H1/TO promoters of varying length and a U6/TO promoter can edit DNA with similar efficiency in vitro, in a Dox dependent manner.
H1/TO promoters of varying length and a U6/TO promoter can edit DNA with similar efficiency in vitro in a doxycycline-dependent manner.
We show that H1/TO promoters of varying length and a U6/TO promoter can edit DNA with similar efficiency in vitro, in a Dox dependent manner.
H1/TO promoters of varying length and a U6/TO promoter can edit DNA with similar efficiency in vitro in a doxycycline-dependent manner.
We show that H1/TO promoters of varying length and a U6/TO promoter can edit DNA with similar efficiency in vitro, in a Dox dependent manner.
The system might be cross compatible with many existing S. pyogenes Cas9 systems and could potentially render those systems inducible.
This system might be cross compatible with many existing S. pyogenes Cas9 systems (i.e., Cas9 mouse, CRISPRi, etc.), and therefore it likely can be used to render these systems inducible as well.
The system might be cross compatible with many existing S. pyogenes Cas9 systems and could potentially render those systems inducible.
This system might be cross compatible with many existing S. pyogenes Cas9 systems (i.e., Cas9 mouse, CRISPRi, etc.), and therefore it likely can be used to render these systems inducible as well.
The system might be cross compatible with many existing S. pyogenes Cas9 systems and could potentially render those systems inducible.
This system might be cross compatible with many existing S. pyogenes Cas9 systems (i.e., Cas9 mouse, CRISPRi, etc.), and therefore it likely can be used to render these systems inducible as well.
The system might be cross compatible with many existing S. pyogenes Cas9 systems and could potentially render those systems inducible.
This system might be cross compatible with many existing S. pyogenes Cas9 systems (i.e., Cas9 mouse, CRISPRi, etc.), and therefore it likely can be used to render these systems inducible as well.
The system might be cross compatible with many existing S. pyogenes Cas9 systems and could potentially render those systems inducible.
This system might be cross compatible with many existing S. pyogenes Cas9 systems (i.e., Cas9 mouse, CRISPRi, etc.), and therefore it likely can be used to render these systems inducible as well.
The system might be cross compatible with many existing S. pyogenes Cas9 systems and could potentially render those systems inducible.
This system might be cross compatible with many existing S. pyogenes Cas9 systems (i.e., Cas9 mouse, CRISPRi, etc.), and therefore it likely can be used to render these systems inducible as well.
The system might be cross compatible with many existing S. pyogenes Cas9 systems and could potentially render those systems inducible.
This system might be cross compatible with many existing S. pyogenes Cas9 systems (i.e., Cas9 mouse, CRISPRi, etc.), and therefore it likely can be used to render these systems inducible as well.
The inducible gRNAi AAV vector is designed to express gRNA from an H1/TO promoter and to express TetR for doxycycline-dependent regulation of gRNA expression.
Specifically, we developed an inducible gRNA (gRNAi) AAV vector that is designed to express the gRNA from a H1/TO promoter. This AAV vector is also designed to express the Tet repressor (TetR) to regulate the expression of the gRNAi in a Dox dependent manner.
The inducible gRNAi AAV vector is designed to express gRNA from an H1/TO promoter and to express TetR for doxycycline-dependent regulation of gRNA expression.
Specifically, we developed an inducible gRNA (gRNAi) AAV vector that is designed to express the gRNA from a H1/TO promoter. This AAV vector is also designed to express the Tet repressor (TetR) to regulate the expression of the gRNAi in a Dox dependent manner.
The inducible gRNAi AAV vector is designed to express gRNA from an H1/TO promoter and to express TetR for doxycycline-dependent regulation of gRNA expression.
Specifically, we developed an inducible gRNA (gRNAi) AAV vector that is designed to express the gRNA from a H1/TO promoter. This AAV vector is also designed to express the Tet repressor (TetR) to regulate the expression of the gRNAi in a Dox dependent manner.
The inducible gRNAi AAV vector is designed to express gRNA from an H1/TO promoter and to express TetR for doxycycline-dependent regulation of gRNA expression.
Specifically, we developed an inducible gRNA (gRNAi) AAV vector that is designed to express the gRNA from a H1/TO promoter. This AAV vector is also designed to express the Tet repressor (TetR) to regulate the expression of the gRNAi in a Dox dependent manner.
The inducible gRNAi AAV vector is designed to express gRNA from an H1/TO promoter and to express TetR for doxycycline-dependent regulation of gRNA expression.
Specifically, we developed an inducible gRNA (gRNAi) AAV vector that is designed to express the gRNA from a H1/TO promoter. This AAV vector is also designed to express the Tet repressor (TetR) to regulate the expression of the gRNAi in a Dox dependent manner.
The inducible gRNAi AAV vector is designed to express gRNA from an H1/TO promoter and to express TetR for doxycycline-dependent regulation of gRNA expression.
Specifically, we developed an inducible gRNA (gRNAi) AAV vector that is designed to express the gRNA from a H1/TO promoter. This AAV vector is also designed to express the Tet repressor (TetR) to regulate the expression of the gRNAi in a Dox dependent manner.
The inducible gRNAi AAV vector is designed to express gRNA from an H1/TO promoter and to express TetR for doxycycline-dependent regulation of gRNA expression.
Specifically, we developed an inducible gRNA (gRNAi) AAV vector that is designed to express the gRNA from a H1/TO promoter. This AAV vector is also designed to express the Tet repressor (TetR) to regulate the expression of the gRNAi in a Dox dependent manner.
In vivo genome editing can be induced with this system by supplying animals doxycycline-containing food for as little as 1 day.
Genome editing can be induced in vivo with this system by supplying animals Dox containing food for as little as 1 day.
minimum induction duration 1 day
In vivo genome editing can be induced with this system by supplying animals doxycycline-containing food for as little as 1 day.
Genome editing can be induced in vivo with this system by supplying animals Dox containing food for as little as 1 day.
minimum induction duration 1 day
In vivo genome editing can be induced with this system by supplying animals doxycycline-containing food for as little as 1 day.
Genome editing can be induced in vivo with this system by supplying animals Dox containing food for as little as 1 day.
minimum induction duration 1 day
In vivo genome editing can be induced with this system by supplying animals doxycycline-containing food for as little as 1 day.
Genome editing can be induced in vivo with this system by supplying animals Dox containing food for as little as 1 day.
minimum induction duration 1 day
In vivo genome editing can be induced with this system by supplying animals doxycycline-containing food for as little as 1 day.
Genome editing can be induced in vivo with this system by supplying animals Dox containing food for as little as 1 day.
minimum induction duration 1 day
In vivo genome editing can be induced with this system by supplying animals doxycycline-containing food for as little as 1 day.
Genome editing can be induced in vivo with this system by supplying animals Dox containing food for as little as 1 day.
minimum induction duration 1 day
In vivo genome editing can be induced with this system by supplying animals doxycycline-containing food for as little as 1 day.
Genome editing can be induced in vivo with this system by supplying animals Dox containing food for as little as 1 day.
minimum induction duration 1 day
The inducible gRNAi vector can edit neuronal genomes in the mouse brain in vivo in a doxycycline-dependent manner.
We also demonstrate that our inducible gRNAi vector can be used to edit the genomes of neurons in vivo within the mouse brain in a Dox dependent manner.
The inducible gRNAi vector can edit neuronal genomes in the mouse brain in vivo in a doxycycline-dependent manner.
We also demonstrate that our inducible gRNAi vector can be used to edit the genomes of neurons in vivo within the mouse brain in a Dox dependent manner.
The inducible gRNAi vector can edit neuronal genomes in the mouse brain in vivo in a doxycycline-dependent manner.
We also demonstrate that our inducible gRNAi vector can be used to edit the genomes of neurons in vivo within the mouse brain in a Dox dependent manner.
The inducible gRNAi vector can edit neuronal genomes in the mouse brain in vivo in a doxycycline-dependent manner.
We also demonstrate that our inducible gRNAi vector can be used to edit the genomes of neurons in vivo within the mouse brain in a Dox dependent manner.
The inducible gRNAi vector can edit neuronal genomes in the mouse brain in vivo in a doxycycline-dependent manner.
We also demonstrate that our inducible gRNAi vector can be used to edit the genomes of neurons in vivo within the mouse brain in a Dox dependent manner.
The inducible gRNAi vector can edit neuronal genomes in the mouse brain in vivo in a doxycycline-dependent manner.
We also demonstrate that our inducible gRNAi vector can be used to edit the genomes of neurons in vivo within the mouse brain in a Dox dependent manner.
The inducible gRNAi vector can edit neuronal genomes in the mouse brain in vivo in a doxycycline-dependent manner.
We also demonstrate that our inducible gRNAi vector can be used to edit the genomes of neurons in vivo within the mouse brain in a Dox dependent manner.
The paper reports development of a viral-mediated CRISPR/Cas9 system with doxycycline-inducible gRNA expression that can be delivered by AAV in vitro and in vivo.
Here, we report the development of a viral mediated CRISPR/Cas9 system that can be rendered inducible utilizing doxycycline (Dox) and can be delivered to cells in vitro and in vivo utilizing adeno-associated virus (AAV).
The paper reports development of a viral-mediated CRISPR/Cas9 system with doxycycline-inducible gRNA expression that can be delivered by AAV in vitro and in vivo.
Here, we report the development of a viral mediated CRISPR/Cas9 system that can be rendered inducible utilizing doxycycline (Dox) and can be delivered to cells in vitro and in vivo utilizing adeno-associated virus (AAV).
The paper reports development of a viral-mediated CRISPR/Cas9 system with doxycycline-inducible gRNA expression that can be delivered by AAV in vitro and in vivo.
Here, we report the development of a viral mediated CRISPR/Cas9 system that can be rendered inducible utilizing doxycycline (Dox) and can be delivered to cells in vitro and in vivo utilizing adeno-associated virus (AAV).
The paper reports development of a viral-mediated CRISPR/Cas9 system with doxycycline-inducible gRNA expression that can be delivered by AAV in vitro and in vivo.
Here, we report the development of a viral mediated CRISPR/Cas9 system that can be rendered inducible utilizing doxycycline (Dox) and can be delivered to cells in vitro and in vivo utilizing adeno-associated virus (AAV).
The paper reports development of a viral-mediated CRISPR/Cas9 system with doxycycline-inducible gRNA expression that can be delivered by AAV in vitro and in vivo.
Here, we report the development of a viral mediated CRISPR/Cas9 system that can be rendered inducible utilizing doxycycline (Dox) and can be delivered to cells in vitro and in vivo utilizing adeno-associated virus (AAV).
The paper reports development of a viral-mediated CRISPR/Cas9 system with doxycycline-inducible gRNA expression that can be delivered by AAV in vitro and in vivo.
Here, we report the development of a viral mediated CRISPR/Cas9 system that can be rendered inducible utilizing doxycycline (Dox) and can be delivered to cells in vitro and in vivo utilizing adeno-associated virus (AAV).
The paper reports development of a viral-mediated CRISPR/Cas9 system with doxycycline-inducible gRNA expression that can be delivered by AAV in vitro and in vivo.
Here, we report the development of a viral mediated CRISPR/Cas9 system that can be rendered inducible utilizing doxycycline (Dox) and can be delivered to cells in vitro and in vivo utilizing adeno-associated virus (AAV).
Approval Evidence
1 source2 linked approval claimsfirst-pass slug h1-to-promoter
We show that H1/TO promoters of varying length and a U6/TO promoter can edit DNA with similar efficiency in vitro, in a Dox dependent manner.
Source:
comparative performancesupports
H1/TO promoters of varying length and a U6/TO promoter can edit DNA with similar efficiency in vitro in a doxycycline-dependent manner.
We show that H1/TO promoters of varying length and a U6/TO promoter can edit DNA with similar efficiency in vitro, in a Dox dependent manner.
Source:
design featuresupports
The inducible gRNAi AAV vector is designed to express gRNA from an H1/TO promoter and to express TetR for doxycycline-dependent regulation of gRNA expression.
Specifically, we developed an inducible gRNA (gRNAi) AAV vector that is designed to express the gRNA from a H1/TO promoter. This AAV vector is also designed to express the Tet repressor (TetR) to regulate the expression of the gRNAi in a Dox dependent manner.
Source:
Comparisons
Source-backed strengths
In the cited work, H1/TO promoter variants of different lengths supported doxycycline-dependent DNA editing in vitro with efficiency similar to a U6/TO promoter. The promoter was also incorporated into an inducible gRNA AAV vector design together with TetR, indicating suitability for viral vector-based regulatory circuits.
Source:
We show that H1/TO promoters of varying length and a U6/TO promoter can edit DNA with similar efficiency in vitro, in a Dox dependent manner.
Ranked Citations
- 1.