A brief introduction about lentiviral vector gene delivery system and its application in CAR-T cell construction. Creative Biolabs offers high quality lentivirus based CAR gene delivery service to help with your CAR-T cell development projects.
2. VIRAL VECTORS——BIOLOGICAL TOOLS FOR GENE DELIVERY
Virus infect target cell
Inject gene into target cell plasma
Viral gene integrate into host genome (retroviral)
Viral gene expression
• Use viral as vector to deliver exogenous genes into target cell
2
3. • Adenoviral vectors
• Adeno-associated vectors
• Retroviral vector
• Lentiviral vector
• Poxvirus vector
DIFFERENT VIRAL VECTOR SYSTEMS 3
• Different virus are used as vectors for gene delivery
• Each system has its own pros and cons
• Always choose the most appropriate system to optimize gene delivery results
4. Gene
insert
size
Transgene
expression
time
Immuno
genicity
Safety Advantages Disadvantages
Adenoviral
vectors
~30kb Short(days) High Low
• high virus titers;
• High level gene expression;
• large insert capacity;
• infects dividing and non-dividing cells.
• Immune response to viral proteins;
• no integration into host genome;
• transient gene expression.
Adeno-associate
d vectors
~4kb
Long(months,
years, maybe life
long)
low high
• Infect dividing and non-dividing cells;
• broad cell tropism;
• potential of targeted integration;
• low immunogenicity.
• Limited capacity for transgenes;
• difficult generation of high virus
titers;
Retroviral vector ~10kb Life long low
Unacceptably
low
• Stable integration into host DNA;
• broad cell tropism of infectivity;
• relatively easy manipulation
• Difficult targeting of viral infection;
• no infection of nondividing cells
• random integration
• instability of vectors
Lentiviral vector ~10kb Life long Low acceptable
• Infect dividing and non-dividing cells
• stable gene expression,
• insert capacity of 10kb
• Potential insertional mutagenesis
• accessory protein sequences in
the packaging constructs
Poxvirus vector ~20kb Short(days) High Relatively high
• High insertion capacity
• high transgene expression level
• Potential cytopathic effects
Retroviral and lentiviral vectors are two of the most commonly used viral vector systems
DIFFERENT VIRAL VECTOR SYSTEMS 4
5. WHY CHOOSE LENTIVIRAL VECTOR 5
One of the best
choice as tools for T
cell gene editing
Loaded gene length as
large as 9kb
Transgene integrate into host
genome, stable expression
T cell as nature
hosts
Both dividing and non-dividing cell
can be infect
Broad tissue tropism
Low immunogenicity
6. 6
long terminal repeat.
regulatory proteins, which activate viral transcription and
control the splicing and nuclear exports of viral transcripts
respectively.
Encodes gp160 protein and cleaved into gp120(SU)
and gp41(TM)
Encodes three essential replication enzymes: RT, IN and PR.
Tat and rev:
Encodes Matrix(MA), capsid protein (CA), nucleocapsid protein (NC)
and p6.
Env:
Pol:
Gag:
LTR:
BASIC KNOWLEDGE ABOUT LENTIVIRUS——STRUCTURE AND GENOME
8. Transfer plasmid Packaging plasmidEnvelope plasmid
2nd generation lentiviral gene delivery system
HOW DOES THE LENTIVIRAL VECTOR SYSTEM WORK? 8
9. • Lentiviral vectors are preferred by oncologists for the generation of CAR-T cells :
LENTIVIRAL VECTOR IN CAR-T DEVELOPMENT 9
Stable transduction efficiency
Low vector related genotoxicity
Available for Scale-up production
Low cost
10. • CAR Construction Kit
• CAR Virus Packing Kit
• CAR Transfection Kit
At Creative Biolabs, our scientists are able to conduct versatile cell
transfection in multiple cell lines, primary cells, stem cells,
hematopoietic cells, and neuronal cells. Creative Biolabs offers the most
professional CAR transfection in T cells and NK cells for the most
cutting-edge genetically modified immunotherapies. The customized
service plans are also welcome since we could offer every service and
experiment kit for every step, including plasmid construction, virus
packaging, cell transfection, etc.
10WHERE CAN I GET GENE DELIVERY SERVICE FOR CAR T DEVELOPMENT?
11. 45-1 Ramsey Road, Shirley, NY 11967, USA
1-631-871-5806 1-631-614-7828
info@creative-biolabs.com www.creative-biolabs.com/car-t/
CONTACT US
LTR: long terminal repeat. subdivided in three functional regions: the U3 (HIV promoter), the R and U5 regions, involved in RNA replication and transcription.
LTR: long terminal repeat. subdivided in three functional regions: the U3 (HIV promoter), the R and U5 regions, involved in RNA replication and transcription.