Functional Genomics & RNAi

Lentiviral Packaging Mix

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The MISSION® Lentiviral Packaging Mix is an optimized formulation for enabling the generation of high-titer lentivirus in HEK293T cells.

  • Accelerate viral production by skipping tedious but critical multivariable plasmid optimization.

  • Produce high titer lentivirus with MISSION shRNA plasmids with the included easy to use protocol.

  • Safely produce lentivirus utilizing the three plasmid system with self inactivating, long terminal repeat (SIN/LTR) regions.

  • Choose between small or large scale lentivirus production with our conveniently sized 250 µl or 1.7 ml sizes.

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Product Overview
Comparison of Viral Delivery Systems
Product Listing

Product Overview

The plasmids included in the lentiviral packaging mix, Figure 1, encode the key structural viral packaging genes and a heterologous viral envelope gene. When combined with a MISSION shRNA pLKO.1 plasmid, this product yields effective high-titer lentivirus, Table 1. Viruses are uniquely adapted to infect even the most resilient cell lines. Therefore, viral systems have been genetically engineered over the last few decades to safely deliver transgenes of interest to target cells. While each viral system has features that are ideal in certain applications, the lentiviral system is an example of a specific retroviral system that has undergone multiple generations of engineering and modification, see Table 2. Generating viral particles can be challenging. We highly recommend the viral format of MISSION TRC shRNA clones to new and experienced users. Small and large scale generation of lentiviral particles has been perfected by Sigma®.


Packaging Vector
Transfer Vector Envelope Vector

Figure 1. For enhanced safety, the structural and replication genes necessary to produce viral particles are separated onto multiple plasmids. All wild-type virulence and accessory genes are deleted. MISSION TRC transfer vectors contain a modified, self-inactivating 3' long terminal repeat (SIN/LTR) which renders the resulting lentiviral particles replication incompetent. Lentiviral particles are packaged in producer cell lines such as HEK293T cells. Upon co-transfection of the plasmids, all required sequences are available to produce and package a viral particle containing the transgene of interest. Only the region between the viral LTRs of the transfer vector is packaged within the viral capsid.



Table 1. Features of the MISSION Lentiviral Packaging System

Feature Result
Multi-plasmid approach No single plasmid contains all the genes necessary to produce packaged lentivirus. Resultant particles are replication-incompetent.
Deletion in U3 portion of 3' LTR which eliminates the promoter-enhancer region Avoids promoter interference issues and further negates the possibility of viral replication.
Elimination of the majority of lentiviral genes (? vpr, vif, vpu and nef) Removes virulence genes which are not necessary for shRNA packaging systems.

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Table 2. Comparison of Viral Delivery Systems

Feature Adenovirus AAV* Retrovirus Lentivirus
Infects non-dividing cells X X   X
Stable integration into host genome   X X X
Can be pseudotyped   X X X
Accepts large inserts X   X X
Lack of interferon response     X X
No issue with transcriptional silencing within genome N/A     X
*Adeno-Associated Virus

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Product Listing

Product No. Product Name Reactions
96-well plate
Reactions
60 mm dish
Add to Cart
SHP001 - 0.25 ml MISSION Lentiviral Packaging Mix 250 25
SHP001 - 1.70 ml MISSION Lentiviral Packaging Mix 1700 170

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Contact Us
For questions about the library, pricing and quotes or other concerns, please e-mail us at: MISSIONRNAi@sial.com.



MISSION is a registered trademark of Sigma-Aldrich Co. LLC Label License.

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