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Lentivirus Packaging

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Lentivirus Packaging
We use 3rd generation lentiviral systems for the highest biosafety standards. Several additional viral sequences must be acquired from the packaging plasmids via recombination.
◆ Description
  • Lentiviral transfer plasmid encoding your insert of interest: The transgene sequence is flanked by long terminal repeat (LTR) sequences, which facilitates integration of the transfer plasmid sequences into the host genome. Typically, it is the sequences between and including the LTRs that is integrated into the host genome upon viral transduction. For safety reasons, transfer plasmids are all replication incompetent and may contain an additional deletion in the 3'LTR, rendering the virus “self-inactivating” (SIN) after integration.
  • Packaging plasmids: One encoding Rev and one encoding Gag and Pol.
  • Envelop plasmid.

Self-inactivating VSV-G pseudotyped lentivirus

 

◆ Lentivirus Production

Transfect 293T packaging cells with transfer plasmid, envelope plasmid and packaging plasmids. Remove media, replace with fresh media. After 48~72 hours, harvest virus. Harvested virus can be used in vitro study. After further purification, ultra-purified lentivirus can be used in vivo study.

 

◆ Quality Control for Virus

Lentiviral titer test will be conducted to make sure only qualified lentivirus entering into your laboratory. For luciferase reporter gene labeled lentivirus, we will perform viral transduction assay and further confirm the titer by qPCR.

 

◆ Service
Lentivirus Packaging Deliverable Turnaraound Application
Standard-scale Lentivirus
(Titer: ≥1×108 TU/ml, Volume: 1ml)
2-3 weeks Cell transduction
Large-scale Lentivirus 
(Titer: 
≥1×109 TU/ml, Volume: 1ml)
2-3 weeks Cell transduction

Cyagen can also provide custom shRNA and gRNA lentivirus packaging services.

 

◆ CRISPR/Cas9 Gene Knockout Lentiviral Vector Construction

The CRISPR/Cas9 system can be used for knocking out gene expression in vivo or in vitro. By using lentiviral-based CRISPR constructs, expressed gRNA and Cas9 can be used in dividing or non-dividing cells or whole model organisms.

Two vector CRISPR-Cas9 system: Lentiviral gRNA vector and Lentiviral Cas9 vector.

 

1. Lentiviral-based gRNA Vector

Lentiviral gRNA vector consists of lentiviral genome sequence from 5’ LTR to 3’ LTR and bacterial plasmid sequence which contains an Ampicillin selection cassette and a pUC origin of replication.

 

2. Lentiviral-based Cas9 Vector

CBh promoter drives co-expression of spCas9 and resistance gene.

For two vector CRISPR-Cas9 system, two different drug selection cassettes will be used to select gRNA and Cas9 co-expressed clones.

 

◆ Quality Control for Vector

We will perform sequencing assay as well as restriction enzyme analysis for every project to make sure only qualified vectors enter your laboratory.

 

◆ Inquiries and Quote Requests

Request a quote now. Alternatively, you can always email cell-service@cyagen.com to inquire about our services or obtain a quote for your project.

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