Adenovirus

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

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 B AC 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

Engineered Adenovirus vectors for transient and quantitative gene expression or knockdown

Benefits

Custom constitutive

Custom inducible

High titer for in vivo

Premade AVs