Different virus lines are better for some applications than others. Choosing the best virus to meet your experimental goals can be difficult, but we are here to help. Below is a chart with a general overview of our viral vectors. If you have more detailed questions, please e-mail us at vectors@uiowa.edu or call us at (319) 335-6726 and we would be happy to discuss your experiment and how the Viral Vector Core can help your lab. 

Comparison of Vectors at a Glance

*Replication defective.
** Replication defective in mammalian cells. Replication competent in insect cells

Replication defective means that critical portions of the viral genome have been removed such that the viral vector can no longer replicate. The removal of these genes makes room for insertion of the genetic material that the investigator wants transduced into target cells. For production of viral vectors that can transduce target cells, the critical viral genes are expressed in trans in packaging cells. The particles that are generated from the packaging cells contain the recombinant replication defective genome.

The nature of the deletions that make the various vectors replication defective are explained under the description for each vector.

A literature search is the best place to start. Pilot studies may be needed to test various serotypes in your experimental system. We offer a serotype testing kit with the CMVeGFP control package with serotypes: AAV2/1, AAV2/2, AAV2/4, AAV2/5, AAV2/6, AAV2/8, AAV2/9, AAV2/rH10, AAV2/PHP.B, AAV2/PHP.eB and AAV2/PHP.S. See the In-Stock Vector page. Below is a list of tropisms for various serotypes based on current literature.  

Any previously produced custom virus can be ordered in iLabs by going to the Request Services tab and choosing Re-order Custom Vectors. You do not need to initiate a new construct project. If it has been many years since we last made the virus, we may ask you to provide the sequence and map of the construct to update our information in iLabs. We often need this information to maxi prep the plasmid and confirm the integrity by digest or to titer the virus by QPCR. We require a minimum purchase of at least 1ml for Adenovirus and 500ul for AAV or Lentiviral vectors to offset the cost of production.  Note: all prices are subject to change depending on customer type-contact us for a more accurate quote at Vectors@uiowa.edu.

Different virus lines have different final buffers, please inquire for details.

Titers are determined differently for each vector type and may use a combination of methods to assess the total number of particles (both live and dead/empty) or the total number of live “infectious” particles. See the details below for the standard titer units reported and the range expected for each vector.

Adenovirus titers are reported as an “infectious” or transduced titer in plaque forming units per ml (pfu/ml) determined by an immunostain of the Adenovirus hexon protein in transduced 293 cells. The concentration of viral DNA and protein is also measured in purified adenoviral preps at OD260 to determine the total number of recombinant adenovirus particles (pt/ml) and assess the ratio of “infectious” to “non-infectious” particles.

 Helper Dependent Adenovirus titers are given as an “infectious" or transduced titer in IGU/ml (Infectious Genomic Units/ml) determined by ddPCR of genomic DNA extracted from transduced cells. HDAd vectors are also assessed for transduced titer by FACs when fluorescent reporters are available; this titer is available upon request.

AAV titers are given as a “physical” titer in viral genomes per ml (vg/ml) determined by direct ddPCR of purified vector particles. Vectors are also assessed for transduced titer by FACs when fluorescent reporters are available. Due to serotype complications, not all transduced titers are comparable so they are therefore reported only when requested. All AAV titers are further confirmed by Silver Stain to check that the staining density agrees with the total particles loaded based on the vg/ml and to confirm the packaging envelope proteins are in the correct/functional ratios.

Lentiviral titers are given as an “infectious” or transduced titer in TU/ml (Transducing Units/ml) determined by FACS analysis of transduced HT1080 cells when fluorescent reporters are available. When this is not possible, lentiviral vector titers are determined by ddPCR of genomic DNA extracted from transduced HT1080 cells. The result is reported in TU/ml after normalization to a GFP-expressing vector.

Vaccinia titers are reported as an "infectious" or transduced titer in plaque forming units per ml (pfu/ml) determined by immunostain of a Vaccinia surface protein in transduced cells.

The vector core makes every effort to provide low endotoxin, sterile vectors suitable for research work in vitro and in vivo. Infectious titers are measured whenever possible to guarantee quality, functional virus is being produced. In addition, the Viral Vector Core checks internal catalog vectors for Luciferase and Cre Recombinase expression.

Ad5 Vectors: Quality control assays include a transduced titer in plaque forming units per ml, a physical measure of particles per mL, and an assay for replication competent adenovirus (RCA) contamination.

HDAd Vectors: Quality control assays include a transduced titer in plaque forming units per ml, a physical measure of particles per mL, an assay for replication competent adenovirus (RCA) and helper virus contamination, and FACs when appropriate.

AAV Vectors: Quality control assays include a physical titer (viral genomes/ml) by ddPCR, transduced titer by FACs when appropriate, and silver stain to further assess virus titer and packaging efficiency.

Lentiviral Vectors: Quality control assays include a transduced titer by ddPCR and transduced titer by FACs when appropriate.

Vaccinia Vectors: Quality control assays include a transduced titer in plaque forming units per ml.

We require gene information, a sequence, map, and recent sequence confirmation upon submission of all new projects. Every effort is made to deliver a high titer, high quality viral vector. All plasmids must be recently sequenced. Despite best efforts, some genes of interest may confer cellular toxicity that results in lower vector titers. If problems are noted, we will work with investigators on a case by case basis to troubleshoot.

Ad5 and HDAd Plasmid Quality Control

One of the main rate limiting steps in Ad5 production is the quality of DNA for the initial transfection and recombination. We highly recommend that you use a good quality endotoxin free maxi prep. We can provide this optional service.

AAV Plasmid Quality Control

Because purifications in AAV may be greatly influenced by sequence, preps will not be repeated and charges will apply to the first prep regardless of outcome in the following situations:

• Constructs that are over packaging capacity. This is ~4.7 kb from ITR to ITR for single stranded AAV and ~2.2 kb from ITR to ITR for self-complementary AAV.
• Where the shuttle backbone size is equal to or smaller than the ITR to ITR transgene. AAV ITRs are bi-directional and may take the path of least resistance where the shuttle backbone is very small as compared to the transgene cassette leading to inappropriate packaging of the shuttle backbone. Reports have also been published of the inappropriate packaging of the Rep or Cap Sequences. J Virol. Jan 2003; 77(1): 776–781.Salvetti et al. Our testing indicates this is more common with small backbone AAV plasmids.
• Involve a novel or modified capsid.

​If problems do arise, investigators will be notified and we will discuss the pros, cons, and finances of proceeding with the project. Sometimes the gamble to proceed with plasmids that fall under these criteria are warranted and we will do our best to guide you in the decision process.

Lenti Plasmid Quality Control

One of the main rate limiting steps in Lentivirus production is the quality of DNA for the initial transfection and recombination. We highly recommend that you use a good quality endotoxin free maxi prep. We can provide this optional service.

Replication competent Adenoviruses (RCAs) are wild-type like viruses in a population of replication-deficient viruses. RCAs result from a crossover event where the deleted E1 region of the virus re-inserts itself into the virus while being grown in packaging cells. Once this happens, the RCA adenovirus will replicate without the need of a packaging cell line and outgrow recombinant adenovirus in packaging cells. To avoid the occurrence of RCA, viruses should not be serially propagated. Recombinant adenovirus with high RCA can be rescued by clonal isolation after plaque assay. 

All investigator plasmids, sequences, and viruses are kept private and confidential. This is always our number one priority. However, investigators may or may not choose to allow us to give out their contact information for the purposes of inquiry and granting access to their vectors. Investigators also have the option of choosing to give broad or specific permission for certain vectors to other parties. In all instances, vectors will never be publically listed or made available to anyone else without specifically written, direct permission to do so which may likely involve a Material Transfer Agreement. 

Some Adenovirus and HDAd catalog vectors are stored in 1 mL aliquots, but some vectors may be available in small aliquots (100uL).

AAV control stocks are available in 25 μL aliquots.

Lenti control stocks are available in 100 μL aliquots only.

For MMLV, Vaccinia and Baculovirus stocks, please inquire.

Investigator Viruses

All vector lines maintain an inventory aliquot of the original plasmid submitted from the investigator for vector construction and can reproduce the vector from this plasmid stock. In addition, Adenoviral and HDAd vectors maintain a glycerol stock of the viral vector that can be amplified more quickly than having to start from the plasmid recombination step. Similarly, Adeno-associated viral vectors produced by the Baculovirus system are maintained long term as a P1 or P2 baculovirus stock that can be readily amplified to AAV vector without repeating the transposition, transfection, and amplification steps.

All of our vectors are titered after 1 freeze thaw cycle so that the titer will be equivalent to what you will use in your first experiment. We have tested the stability of vectors through multiple freeze and thaw cycles and find that the titer is consistent for approximately 3 freeze and thaw cycles before it begins to drop; however, we recommend that investigators limit freeze/thaw cycles to as few as possible. Vectors are fairly stable for short periods of 24-48 hours at 4 degrees. Storage instructions can be found on the Technical Resources page. We recommend that the vector be stored at -80 upon receipt. Thaw the vector only when you are ready to use it and make aliquots of no less than 25 μl in 0.5 ml tubes.