Source 1primary paper2016Methods in molecular biology
Toolkit/adenoviral infection
adenoviral infection
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Adenoviral infection is a viral delivery harness used in vitro to introduce the optogenetic actuators ChR2(H134R) and ArchT into primary cardiac fibroblasts. In the cited Methods in Molecular Biology protocol, it enables quick, robust, and consistent opsin expression and supports generation of light-responsive cardiac fibroblast preparations.
Usefulness & Problems
Why this is useful
This delivery approach is useful for establishing optogenetically modified primary cardiac fibroblasts for in vitro studies. The cited source indicates that these modified fibroblasts can be co-cultured with non-transformed cardiomyocytes to obtain a light-sensitive excitable cardiac syncytium.
Problem solved
It addresses the practical need to achieve reliable expression of optogenetic tools in primary cardiac fibroblasts. This is specifically framed as enabling optical actuation of otherwise non-excitable cardiac cells in vitro.
Published Workflows
Objective: Introduce optogenetic actuation tools into primary cardiac fibroblasts, create hybrid co-cultures with cardiomyocytes to obtain light-sensitive excitable cardiac syncytium, and functionally test opsin responsiveness for mechanistic study of cardiac bioelectric coupling.
Why it works: The abstract presents a sequence in which adenoviral delivery yields quick, robust, and consistent opsin expression in primary cardiac fibroblasts, enabling their incorporation into co-cultures and subsequent all-optical testing of opsin responsiveness.
optogenetic actuation of non-excitable cardiac fibroblastsbioelectric contribution of non-excitable heart cellselectrical coupling to cardiomyocytesadenoviral infectionhybrid co-culturestencil-patterned cell growthall-optical functional testing
Stages
- 1.Opsin delivery into primary cardiac fibroblasts(library_build)
This stage establishes optogenetically modified cardiac fibroblasts as the substrate for downstream co-culture and functional testing.
Selection: successful introduction of ChR2(H134R) and ArchT into primary cardiac fibroblasts by adenoviral infection
- 2.Hybrid co-culture setup with cardiomyocytes(functional_characterization)
This stage creates the hybrid cellular system needed to study how modified fibroblasts contribute to excitable syncytium behavior and coupling to cardiomyocytes.
Selection: specific conditions for co-culture of optogenetically modified cardiac fibroblasts and non-transformed cardiomyocytes
- 3.All-optical functional testing(confirmatory_validation)
This stage functionally tests whether the introduced opsins are responsive after delivery and culture setup.
Selection: responsiveness of the introduced opsins in cardiac fibroblasts
Steps
- 1.Adjust adenoviral infection parametersdelivery method
Tune multiplicity of infection and virus incubation duration for the target lab setting or cell type.
The abstract states that infection parameters are adjusted to generalize the method before or during delivery into primary cardiac fibroblasts.
- 2.Introduce ChR2(H134R) and ArchT into primary cardiac fibroblasts by adenoviral infectionopsin cargo and delivery harness
Generate optogenetically modified primary cardiac fibroblasts.
Opsin expression in fibroblasts is required before co-culture formation and functional testing.
- 3.Create hybrid co-cultures with non-transformed cardiomyocytes
Obtain a light-sensitive excitable cardiac syncytium from optogenetically modified fibroblasts and cardiomyocytes.
Co-culture follows opsin delivery because modified fibroblasts are needed to build the hybrid excitable syncytium.
- 4.Functionally test opsin responsiveness using an all-optical frameworkassay method
Assess responsiveness of the introduced opsins in cardiac fibroblasts.
Functional testing is performed after delivery and culture setup because responsiveness can only be assessed once opsins are expressed in the relevant cellular context.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A delivery strategy grouped with the mechanism branch because it determines how a system is instantiated and deployed in context.
Mechanisms
viral gene deliveryTechniques
No technique tags yet.
Target processes
No target processes tagged yet.
Input: Light
Implementation Constraints
Execution requires adenoviral vectors carrying ChR2(H134R) or ArchT for infection of primary cardiac fibroblasts in vitro. The extraction notes indicate that infection parameters such as multiplicity of infection and virus incubation duration require adjustment, but no further construct or dosing details are provided in the supplied evidence.
The provided evidence is limited to an in vitro protocol context in primary cardiac fibroblasts and co-culture experiments. The source material provided here does not establish in vivo performance, cell-type generality beyond the stated system, or whether optimization can be avoided across preparations.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Specific co-culture conditions of optogenetically modified cardiac fibroblasts with non-transformed cardiomyocytes can be used to obtain a light-sensitive excitable cardiac syncytium.
Specific co-culture conditions of optogenetically modified cardiac fibroblasts with non-transformed cardiomyocytes can be used to obtain a light-sensitive excitable cardiac syncytium.
Specific co-culture conditions of optogenetically modified cardiac fibroblasts with non-transformed cardiomyocytes can be used to obtain a light-sensitive excitable cardiac syncytium.
Specific co-culture conditions of optogenetically modified cardiac fibroblasts with non-transformed cardiomyocytes can be used to obtain a light-sensitive excitable cardiac syncytium.
Specific co-culture conditions of optogenetically modified cardiac fibroblasts with non-transformed cardiomyocytes can be used to obtain a light-sensitive excitable cardiac syncytium.
Specific co-culture conditions of optogenetically modified cardiac fibroblasts with non-transformed cardiomyocytes can be used to obtain a light-sensitive excitable cardiac syncytium.
Specific co-culture conditions of optogenetically modified cardiac fibroblasts with non-transformed cardiomyocytes can be used to obtain a light-sensitive excitable cardiac syncytium.
Specific co-culture conditions of optogenetically modified cardiac fibroblasts with non-transformed cardiomyocytes can be used to obtain a light-sensitive excitable cardiac syncytium.
Specific co-culture conditions of optogenetically modified cardiac fibroblasts with non-transformed cardiomyocytes can be used to obtain a light-sensitive excitable cardiac syncytium.
An all-optical framework is described for functional testing of opsin responsiveness in cardiac fibroblasts.
Adenoviral infection can introduce ChR2(H134R) and ArchT into primary cardiac fibroblasts in vitro and yield quick, robust, and consistent expression.
Adenoviral infection can introduce ChR2(H134R) and ArchT into primary cardiac fibroblasts in vitro and yield quick, robust, and consistent expression.
Adenoviral infection can introduce ChR2(H134R) and ArchT into primary cardiac fibroblasts in vitro and yield quick, robust, and consistent expression.
Adenoviral infection can introduce ChR2(H134R) and ArchT into primary cardiac fibroblasts in vitro and yield quick, robust, and consistent expression.
Adenoviral infection can introduce ChR2(H134R) and ArchT into primary cardiac fibroblasts in vitro and yield quick, robust, and consistent expression.
Adenoviral infection can introduce ChR2(H134R) and ArchT into primary cardiac fibroblasts in vitro and yield quick, robust, and consistent expression.
Adenoviral infection can introduce ChR2(H134R) and ArchT into primary cardiac fibroblasts in vitro and yield quick, robust, and consistent expression.
Adenoviral infection can introduce ChR2(H134R) and ArchT into primary cardiac fibroblasts in vitro and yield quick, robust, and consistent expression.
Adenoviral infection can introduce ChR2(H134R) and ArchT into primary cardiac fibroblasts in vitro and yield quick, robust, and consistent expression.
Adjusting multiplicity of infection and virus incubation duration is presented as a way to generalize the adenoviral infection method across different lab settings or cell types.
Adjusting multiplicity of infection and virus incubation duration is presented as a way to generalize the adenoviral infection method across different lab settings or cell types.
Adjusting multiplicity of infection and virus incubation duration is presented as a way to generalize the adenoviral infection method across different lab settings or cell types.
Adjusting multiplicity of infection and virus incubation duration is presented as a way to generalize the adenoviral infection method across different lab settings or cell types.
Adjusting multiplicity of infection and virus incubation duration is presented as a way to generalize the adenoviral infection method across different lab settings or cell types.
Adjusting multiplicity of infection and virus incubation duration is presented as a way to generalize the adenoviral infection method across different lab settings or cell types.
Adjusting multiplicity of infection and virus incubation duration is presented as a way to generalize the adenoviral infection method across different lab settings or cell types.
Adjusting multiplicity of infection and virus incubation duration is presented as a way to generalize the adenoviral infection method across different lab settings or cell types.
Adjusting multiplicity of infection and virus incubation duration is presented as a way to generalize the adenoviral infection method across different lab settings or cell types.
Approval Evidence
1 source2 linked approval claimsfirst-pass slug adenoviral-infection
introduce optogenetic actuation tools ... into primary cardiac fibroblasts (cFB) in vitro by adenoviral infection to yield quick, robust, and consistent expression
Source:
method capabilitysupports
Adenoviral infection can introduce ChR2(H134R) and ArchT into primary cardiac fibroblasts in vitro and yield quick, robust, and consistent expression.
Source:
method generalizabilitysupports
Adjusting multiplicity of infection and virus incubation duration is presented as a way to generalize the adenoviral infection method across different lab settings or cell types.
Source:
Comparisons
Source-backed strengths
The supplied evidence states that adenoviral infection yields quick, robust, and consistent expression in primary cardiac fibroblasts. It was applied to deliver both ChR2(H134R) and ArchT, supporting preparation of light-responsive fibroblast cultures and co-culture-based light-sensitive cardiac syncytia.
Ranked Citations