A number of self-inactivating lentiviral vectors can be used for transgene delivery and RNAi induction. A list of the vectors is given below.

LentiLox 3.7 (pLL3.7)

LentiLox 3.7 is a lentiviral vector designed for inducing RNA interference in a wide range of cell types, tissues and organisms. We have use this vector to infect and efficiently silence proteins in hematopoetic stem cells and their progeny, and have used infected embryonic stem cells and single cell embryos to create transgenic animals (Rubinson and Dillon et al, Nature Genetics, 2003). Note: Lentiviral vectors have signficant biosafety issues, explained below.

• pLL3.7 Diagram
• pLL3.7 Restriction Map
• pLL3.7 Sequence: Genebank format - Text format

MTA - Licencing issues for plasmid requests

Academic labs can obtain this reagent through ATCC. To order, one should call 1-800-638-6597 and request item VRMC-39.

Item: VRMC-39
Designations: LentiLox 3.7 [pLL3.7]

Industry or biotech laboratories who wish to use this vector must obtain a license throught MIT's Technology Licensing Office.

Note: This vector must be used in conjuction with packaging vectors which are commercially available through Invitrogen (Vira Power Lentiviral Support Kit K4970-00).

Packaging Vectors

We use the 3rd generation packaging systems for lentiviral production originally published by Dull et al. These vectors include: pMDLg/pRRE (gag/pol elements), pRSV-REV, and pMD.G (env elements). Maps are provided courtesy of the Trono laboratory.

• Map of pMDLg/pRRE
• Map of pRSV-REV
• Map of pMD.G

Sources of these packaging vectors include:

• Invitrogen- Viral Power Lentiviral Support Kit K4970-00
• Trono Laboratory, University of Geneva Medical School, Geneva, Switzerland
• Naldini Laboratory, IRCC, University of Torino, Italy - Email
• Verma Laboratory, Salk Institute, San Diego, CA - Email

Biosafety Issues

IMPORTANT: The biosafety office at your institution must be notified prior to use of this system for permission and for further institution-specific instructions. BL2/(+) conditions should be used at all times when handling the virus. All decontamination steps should be performed using 70% ethanol/1% SDS. Gloves should be worn at all times when handling lentiviral preparations, transfected cells or the combined transfection reagent. Just remember that although this virus has been significantly modified for biosafety, it derived from HIV and with a VSV pseudotype human cells can be infected even if they are not dividing. That said, the following modifications have been made to prevent viral replication.

1. Packaging vector lacks both LTRs and has no viral packaging signal (y)
2. The following viral genes have been deleted from the packaging vector: env, tat, rev, vpr, vpu, vif and nef.
3. Rev is supplied in trans on a different vector (RSV-Rev).
4. The vector expressing the packaged viral genome has a self-inactivating LTR (TATA box deletion) and expresses no viral gene products.
5. Envelope, in this case VSVG, is expressed on a separate vector.

For more information please refer to the following papers.

Packaging vectors (pMDLg/pRRE, CMV-VSVG and RSV-Rev):

• Dull et al., A Third-Generation Lentivirus Vector with a Conditional Packaging System. J. Virol. 1998 72(11): 8463-8472.
• Naldini L, Blomer U, Gallay P, Ory D, Mulligan R, Gage FH, Verma IM, Trono D. In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector. Science. 1996 Apr 12;272(5259):263-7.

Self inactivating LTR:

• Miyoshi H, Blomer U, Takahashi M, Gage FH, Verma IM. Development of a self-inactivating lentivirus vector. J Virol. 1998 Oct;72(10):8150-7.