Source 1primary paper2016Frontiers in Molecular Neuroscience
Toolkit/inducible gRNA (gRNAi) AAV vector
inducible gRNA (gRNAi) AAV vector
Also known as: gRNAi AAV vector, inducible gRNAi vector
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The inducible gRNA (gRNAi) AAV vector is an adeno-associated viral construct that places a CRISPR guide RNA under an H1/TO promoter and co-expresses Tet repressor (TetR) for doxycycline-dependent control of gRNA expression. In the cited 2016 study, related H1/TO and U6/TO promoter configurations supported doxycycline-dependent DNA editing in vitro, and the system was described for inducible in vitro and in vivo genome editing.
Usefulness & Problems
Why this is useful
This construct provides inducible control over guide RNA expression within an AAV delivery format, enabling temporal regulation of CRISPR/Cas9 genome editing. The study further suggested that the design might be cross-compatible with existing Streptococcus pyogenes Cas9 systems, which could extend inducibility to established Cas9 platforms.
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 CRISPR guide RNA expression doxycycline responsive rather than constitutive in a viral vector context. This is intended to support inducible genome editing in vitro and in vivo by coupling TetR-mediated repression with doxycycline-triggered derepression.
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 links
Need controllable genome or transcript editing
DerivedThe inducible gRNA (gRNAi) AAV vector is an adeno-associated viral construct that places a CRISPR guide RNA under an H1/TO promoter and co-expresses Tet repressor (TetR) for doxycycline-dependent control of gRNA expression. In the cited 2016 study, related H1/TO and U6/TO promoter configurations supported doxycycline-dependent DNA editing in vitro, and the system was described for inducible in vitro and in vivo genome editing.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A reusable architecture pattern for arranging parts into an engineered system.
Mechanisms
crispr/cas9 genome editingdoxycycline-dependent transcriptional derepressiontetr-mediated repression of grna expressionTechniques
Computational DesignTarget processes
editingImplementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationimplementation constraint: payload burdenoperating role: regulator
The construct is an AAV vector designed to express the gRNA from an H1/TO promoter and to co-express TetR. Doxycycline is required as the inducing small molecule, and the cited work also evaluated H1/TO promoters of varying length and a U6/TO promoter in vitro.
The supplied evidence is limited to one primary study and provides sparse quantitative performance details. Cross-compatibility with many existing S. pyogenes Cas9 systems was presented as a possibility rather than a demonstrated result 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 source4 linked approval claimsfirst-pass slug inducible-grna-grnai-aav-vector
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:
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:
induction timingsupports
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.
Source:
in vivo applicationsupports
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.
Source:
tool developmentsupports
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).
Source:
Comparisons
Source-backed strengths
The design explicitly combines an H1/TO gRNA expression cassette with TetR expression in a single AAV vector for doxycycline-dependent regulation. In vitro, H1/TO promoters of varying length and a U6/TO promoter were reported to edit DNA with similar efficiency in a doxycycline-dependent manner, indicating some promoter-design flexibility.
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.
Compared with IscB cytosine base editors
inducible gRNA (gRNAi) AAV vector and IscB cytosine base editors address a similar problem space because they share editing.
Shared frame: same top-level item type; shared target processes: editing
Compared with NGF-overexpressing mesenchymal stem cells
inducible gRNA (gRNAi) AAV vector and NGF-overexpressing mesenchymal stem cells address a similar problem space because they share editing.
Shared frame: same top-level item type; shared target processes: editing
Compared with synthetic promoters
inducible gRNA (gRNAi) AAV vector and synthetic promoters address a similar problem space because they share editing.
Shared frame: same top-level item type; shared target processes: editing
Relative tradeoffs: appears more independently replicated; looks easier to implement in practice.
Ranked Citations
- 1.