Viral Vector BSDS
Adeno-Associated Virus (AAV)
| Types | 12 serotypes (AAV2 tropism: muscles, neurons, hepatocytes) |
|---|---|
| NIH Risk Group | 1 (AAV 1-4); unless oncogene/toxin or helper virus present (RG-2) |
| Biocontainment Level | BSL-1; unless oncogene/toxin or helper virus present (BSL-2) |
| Infectious to Humans/Animals | Yes (Humans/Primates) Route of Transmission: Inhalation Disease: Not known to cause direct disease in humans; causes mild immune response Treatment: Supportive care |
| Safety Considerations | Infects multiple cell types. May be associated with insertional mutagenesis and cancer. Inserts itself on human chromosome 19 and remains latent. Can be potentially reactivated later in the presence of a helper virus and produce infection. Integration: Selective Replication Compentent: No — needs helper virus (CMV, adenovirus, herpesvirus, vaccinia) RCV Testing: If helper virus is adenovirus, presence of RCV after heat inactivation (56C for 15min) Recombination: Possible. Use of 293 cells with E1 deletion can cause systematic rescue Transgene: Evaluate if oncogenic or biotoxic Ecotropic/Xenotropic/Amphotropic: |
| Safe Use | 1) Gutless systems (Cre-loxP) 2) 2nd Generation: Deletion of regions E1, E2, E3, E4 genes 3) 1st Generation: Deletion of regions E1, E3 genes (less safe) Disinfection: 10% bleach (fresh) |
| Animals | ABSL-1; ABSL-2 in the presence of a helper virus or if host animal could house helper virus. Special handling of bedding and cages for 48 hours post injection. Bedding disposed in biohazardous waste. |
| Vendors | Vector Biolabs, Cell Biolabs |
Adenovirus
| Types | Ad 1, 2, 3, 4, 5, 7, 40, 41 |
|---|---|
| NIH Risk Group | 2 |
| Biocontainment Level | BSL-2. No open bench work. Use eye protection. Stable in environment |
| Infectious to Humans/Animals | Yes Route of Transmission: Oral/Inhalation Disease: Primarily upper respiratory. Generally mild. Can cause eye damage. Treatment: Supportive care |
| Safety Considerations | Can infect a variety of non-dividing cells. Cytotoxic (T-cell response) Integration: No Replication Competent: Possible RCV Testing: Recommended- PCR for E1 prior to use; plate on E1 non-susceptible cell types Recombination: Possible. Use of 293 cells with E1 deletion can cause systematic rescue of wild type Transgene: Evaluate if oncogenic or biotoxic Ecotropic/Xenotropic/Amphotropic: |
| Safe Use |
1) Gutless systems (Cre-loxP 2) 2nd Generation: Deletion of regions E1, E2, E3, E4 genes 3) 1st Generation: Deletion of regions E1, E3 genes (less safe) Disinfection: 10% bleach (fresh); Do not use 70% alcohol. |
| Animals | ABSL-2 housing for 7 days post-injection (replication incompetent); ABSL-2 for xenografts of human/animal cells infected. During post infection period: Animals manipulated in BSC. Microisolators used. Adenovirus is excreted by animals. Cage changes/dumping in BSC. Spray cages/racks with virucide. |
| Vendors | Agilent (AdEasy), MP Biomedicals (AdenoVator), Invitrogen (ViraPower) |
Baculovirus
| Types | Autographa californica | |
|---|---|---|
| NIH Risk Group | 1 | |
| Biocontainment Level | BSL-1 | |
| Infectious to Humans/Animals | Possible with mammalian promoter Route of Transmission: Direct contact Disease: No known disease Treatment: NA |
|
| Safety Considerations | Can be modified to infect mammalian cells. Integration: Yes Replication Competent: No Recombination: Pseudotyped: Possible Transgene: Evaluate if oncogenic or biotoxic Ecotropic/Xenotropic/Amphotropic: Evaluate |
|
| Safe Use | Disinfection: 10% bleach (fresh); 70% ETOH | |
| Animals | ABSL-1 | |
| Vendors |
Retrovirus (Lentivirus)
| Types | Human (HIV 1-3), Human T-cell Lymphotropic Virus Type 4 (HTLV), Simian (SIV), Feline (FIV) | |
|---|---|---|
| NIH Risk Group | 2 or 3 (HIV) | |
| Biocontainment Level | Nonhuman (BSL-2), Human (BSL-2+ or higher) | |
| Infectious to Humans/Animals | Yes Route of Transmission: Bloodborne Disease: Can cause severe immunologic and neurological disease in hosts Treatment: Available for HIV (NRT inhibitors, Protease inhibitors, others) |
|
| Safety Considerations | Insertational mutagenesis. Can infect non-dividing cells including immune cells. Can infect non-target cells. Can persist lifelong. High mutation rates. Inappropriate expression of gene product. Rescue by other human pathogenic viruses Integration: Yes Replication Competent: Possible RCV Testing: Serial transfer and ELISA assay for P24 antigen at 1 infectious unit/mL Recombination: Possible. (Decreases with later generations) Use transient production system Pseudotyped: VSV-G (extends host tropism) Transgene: Evaluate if oncogenic or biotoxic Ecotropic/Xenotropic/Amphotropic: Varies |
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| Safe Use |
1) Use third generation system: recommend multiple plasmid system with Gag, Pol, Env genes on separate plasmids. Risk decreases from two plasmids to four (recombination/replication competent) 2) Tat gene deleted (replication) 3) Use non-native Env gene. Use non-primate vectors if possible. Disinfection: 10% bleach (fresh) |
|
| Animals | Is host permissive? ABSL-2 housing for 48 hours post-injection (replication incompetent); ABSL-2 for xenografts of human/animal cells infected. Safe needle devices for injection. |
|
| Vendors | Invitrogen |
Murine Retroviruses
| Types | Moloney Murine Leukemia Virus: (MoMuLV), Mouse Mammary Tumor Virus (MMTV) |
|---|---|
| NIH Risk Group | 1 (if ecotropic); 2 for others |
| Biocontainment Level | BSL-1 if ecotropic and replication incompetent; BSL-2 for others |
| Infectious to Humans/Animals | Humans: possible if pseudotyped Route of Transmission: Bloodborne Disease: Cell transformation and tumor formation Treatment: |
| Safety Considerations | Insertational mutagenesis possible, leading to cell transformation/tumor formation. Amphotropic Env gene can infect non-murine cells including human cells Integration: Yes Replication Competent: Possible RCV Testing: Use permissive cell line (Mus dunni); screen by marker rescue assay (PG-4S+L-) Recombination: Possible (Helper virus or endogenous virus in cell lines can increase risk) Pseudotyped: Yes Transgene: Evaluate if oncogenic or biotoxic Ecotropic/Xenotropic/Amphotropic: Varies based on packaging cell line |
| Safe Use | 1) Use ecotropic Env gene Disinfection: 10% bleach (fresh) |
| Animals | Depends on construct. ABSL-1 for ecotropic. Virus will be shed in bedding. ABSL-2 if infects humans. |
| Vendors | Invitrogen |
Sindbis (alphavirus/arbovirus)
| Types | |
|---|---|
| NIH Risk Group | 2 |
| Biocontainment Level | BSL-2 (Stable up to two days) |
| Infectious to Humans/Animals | Yes (insects/birds/rodents/humans) Route of Transmission: Bloodborne Disease: Self limiting febrile disease, rash, arthritis Treatment: |
| Safety Considerations | Cytotoxic: virus can induce apoptosis Integration: Replication Competent: RCV Testing: Recombination: Pseudotyped: Transgene: Evaluate if oncogenic or biotoxic Ecotropic/Xenotropic/Amphotropic: |
| Safe Use | Disinfection: 10% bleach (fresh) |
| Animals | ABSL-2 |
| Vendors |
