Engineered Adenovirus vectors for transient and quantitative gene expression or knockdown
SIRION BIOTECH`s patented BAC technology enables adenovirus construction in less than 5 weeks. The system complies with all standard safety criteria by using replication deficient E1/E3 deleted serotype Ad5 as basis. Virtually every desired gene or shRNA expression construct is possible, with a capacity of 7.5 kb.
Benefits
- Transient gene expression in vivo and in vitro
- Large packaging capacity up to 7.5 kb gives a space for multiple gene expression cassettes
- Reliable gene delivery into dividing and non-dividing cells with almost 100% efficiency
- Fast results – gene expression already within 24 h
- High titer productions for transgene delivery in vivo
- Strong ubiquitous and inducible promoters, custom promoters
- Transduction enhancers for hard-to-transduce cell types
- No host genome integration
Any project is thoroughly discussed with our customers to ensure that the resulting particles are feasibly designed and fit the final application.
Custom constitutive
Fields of Application
- Strong, tightly regulated and highly efficient transient gene overexpression and knockdown in vitro
- For primary cells and all hard-to-transfect cell types
- High titer production for in vivo applications
Benefits
- Up to 4 genes from one vector through large packaging capacity (up to 7.5 kb)
- Large promoter and marker gene portfolio
- Highest product purity
- Reproducible results through high-efficiency titration
- Tight control of gene expression levels by adapting virus concentration (MOI)
- knockdown efficiency of at least 80% on mRNA level when combining with our RNAiONE™ service
Service Details
- Cloning of customer cDNA (up to 7.5 kb)/ shRNA in SIRION`s adenovirus shuttle plasmid
- Verification of cloning success by DNA sequencing
- Generation of recombinant adenovirus BACs
- Production of low-passage virus stocks followed by amplification in 293 cells
- Purification, titration of infectious units (IU) and QC
- Gene expression quantification on mRNA level by qRT-PCR in a standard cell system and/or
- Optionally on protein level by Western Blot
- Optionally transient cell model generation
Scale
|
Infectious Units (IU)
|
Time
|
Standard
|
≥ 1 X10E9
|
4-6 weeks
|
Medium
|
≥ 1 X10E10
|
5-8 weeks
|
Large
|
≥ 1 X10E11
|
7-9 weeks
|
X-Large
|
≥ 1-5X10E11
|
9-11 weeks
|
*the table represents timelines for Adenovirus generation, including cloning
Custom inducible
SIRION Biotech’s inducible expression platform is based on the latest 3G TET technology. This guarantees a highly sensitive doxycycline-inducible system without leakiness. Our inducible adenovirus platform is based on a TWO vector system.
Fields of Application:
- Characterization of toxic gene modulations
- Strong, tightly regulated inducible gene overexpression in vitro
- Highly efficient inducible gene knockdown studies in vitro
- For primary cells and hard-to-transfect cell types
Benefits:
- Personal project consultation
- Tightly controlled ON/OFF switch by doxycycline
- Highest product purity
- Reproducible results by high efficient titration
- Tight regulation of gene expression levels with different MOIs
- Large promoter and marker gene portfolio
- Knockdown efficiency of at least 80% on mRNA level when combining with our RNAiONE service
Service Details:
- Cloning of your cDNA (up to 6 kb)/shRNA in SIRION’s inducible adenovirus shuttle plasmid
- Verification of cloning success by DNA sequencing
- Generation of recombinant adenovirus BACs
- Production of low-passage virus stocks followed by amplification in HEK293 cells
- Purification, titration of infectious units (IU) and QC
- Gene expression quantification on mRNA level by qRT-PCR in a standard cell system and/or
- optionally on protein level by Western Blot
- optionally transient inducible cell model generation
Scale
|
Infectious Units (IU)
|
Time
|
Standard
|
≥ 1 X10E9
|
4-6 weeks
|
Medium
|
≥ 1 X10E10
|
5-8 weeks
|
Large
|
≥ 1 X10E11
|
7-9 weeks
|
X-Large
|
≥ 1-5X10E11
|
9-11 weeks
|
*the table shows information for Adenovirus inducible generation, including cloning
Contact us – to discover more
High titer for in vivo
Adenoviruses can be concentrated to high titers of 10E11 IU/ml making them an extremely efficient method of delivering shRNA or transgenes in vivo. Additional benefits for in vivo use are the fast and strong onset of expression. When using adenoviruses, one must take into consideration that >90% of the virus particles will be absorbed by the liver and that there can be strong immunoreactions of the host organism.
An alternative for in vivo injections with versatile and highest tissue tropism, minimal immunogenicity and handling under BL-1 (S1) are AAVs.
Fields of Application
- Efficient transgene/ shRNA expression in vivo
SIRION’s benefits
- Vector formulation for direct in vivo use
- Fast and strong expression onset
- Large promoter and marker gene portfolio
- Flexibility in vector design
- Highly concentrated [5E10 – 5E11 IU/ml]
- From cloning to viral delivery in less than 9 weeks – guaranteed with patented BAC technology
- Up to 4 genes from one vector through large packaging capacity (up to 7.5 kb)
- Highest product purity
- Reproducible results due to standardized product quality
Service Details
- Cloning of your cDNA (up to 7.5 kb)/ shRNA in SIRION`s adenovirus shuttle plasmid
- Verification of cloning success by DNA sequencing
- Generation of recombinant adenovirus BACs
- Production of low passage virus stocks followed by amplification in HEK293 cells
- Upscaling in HEK293 cells to large/ X-large titer
- Specific purification via ultracentrifugation
- Titration of infectious units (IU) and QC
- Gene expression quantification on mRNA level by qRT-PCR in a standard cell system and/ or
- Optionally on protein level by Western Blot
Premade AVs
- Ready to use human Adenovirus Type5 (dE1/E3) with Luc, GFP, CRE
- For transient constitutive overexpression and knockdown
- Validated shRNAs with >80% knockdown guarantee on mRNA level
- Functionality tested
- Titer: 1X10E9 IU [1X10E10 IU/ml]
- Delivery time: 1-2 weeks
