Download Lentiviral Gene Delivery Tools

WORKFLOW
Lentiviral Gene Delivery Tools
Use the typical lentivirus production workflow depicted below to find products, learning
resources, and technical information related to each step of your experiment.
Choose your
vector
Clone your
gene
Package your
virus
Titrate your
virus
Concentrate
your virus
When choosing a plasmid backbone, you have many features to consider for
expression of your gene of interest. Constitutive or inducible expression? Untagged or
fluorescently labeled? Explore available options to find the best vector for your
experimental aim.
In­Fusion cloning is ideal for inserting your fragment into a lentiviral vector—just
one quick reaction allows you to recover your final construct.
Learn how In­Fusion Cloning was used to quickly clone PCR fragments into a
lentiviral vector that had only one suitable restriction site for subcloning.
The most critical factor for successful lentiviral transduction is viral titer. Start with
our fourth generation packaging system and obtain between 107and 108 infectious
units per ml, 25 times what other popular systems generate.
It is important to have an accurate estimation of infectious forming units (IFU) in your
packaging supernatant. This measurement will determine how much supernatant is
used to achieve a desired MOI—and ensure successful gene delivery.
Need a fast answer? Lenti­X GoStix take only 10 minutes to assess lentivirus titer
and determine whether your supernatants are ready for harvesting.
If your IFU/ml titer is low, Lenti­X Concentrator can be used to concentrate your virus
up to 100­fold and improve gene delivery. This concentration method is scalable,
easier, and faster than ultracentrifugation.
Transduction problems can be frustrating and waste valuable resources—inhibitors in
your media or difficult cell types can hinder lentiviral gene delivery. Learn how to
navigate transduction troubles.
Transduce your
target cells
Want to avoid Polybrene and speed up transduction? Bring virus particles from your
lentiviral packaging supernatant and cultured cells together with charged magnetic
beads.
Some cell types, such as hematopoietic cells, show very low transduction efficiency
even with high­titer lentivirus. RetroNectin reagent promotes co­localization of virus
and cells to dramatically enhance transduction efficiency.
Analyze
integration
The life cycle of lentivirus results in integration of a copy of the lentiviral genome into
the host genome. The chosen integration site has important consequences for both
the expression of your transgene and the phenotype of the host cell. Identify the
specific integration site.
http://clontech.com/US/Products/Viral_Transduction/Selection_Guides/Lentiviral_Workflow