Self-inactivating VSV-G pseudotyped lentivirus
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.
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.
(Titer: ≥1×108 TU/ml, Volume: 1ml)
(Titer: ≥1×109 TU/ml, Volume: 1ml)
Cyagen can also provide custom shRNA and gRNA lentivirus packaging services.
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.
We will perform sequencing assay as well as restriction enzyme analysis for every project to make sure only qualified vectors enter your laboratory.