Viral Vectors (La Jolla)

Overview

The goal of the Viral Vector Core Facility is to assist Institute investigators, as well as outside laboratories, with the cost-effective production of high-quality, high-titer lentiviral, retroviral and adenoviral vector. The Viral Vector Core offers a variety of services including ready-to-transduce lentiviral and adenoviral particles, and custom lentiviral and retroviral packaging and titrations. The packaging, concentrating, and tittering of use-supplied lentiviral constructs is the major activity of the core. The more recently implemented “the works” service includes custom lentiviral vector construction for constitutive or inducible cDNA/shRNA expression, viral packaging, and titration (ready-to-transduce control particles provided). In the near future, custom adenoviral productions including amplification, purification, and titration will also be available. The Core is fully equipped for production of consistent quality viral vectors in carefully monitored producer cell lines (293T and 293A). Consultation is provided on how to use the vectors, and to address any other questions.
Tech - Shared Resources: ServicesQuickJump-Contacts and Prices

Services

The core facility focuses on three major types of services:

1. Custom packaging of viral particles from user-supplied constructs – primarily lentiviral vectors, but also packaging of retroviral and adenoviral vectors is now supported. Concentration of viral particles via sucrose gradient ultracentrifugation, and tittering of viral constructs expressing a fluorescent marker is also provided. The core provides the needed packaging plasmids for second or third generation lentiviral vectors, along with the appropriate cell lines, serum-free UltraCULTURE media, and other materials. Viral preps are scalable, typically ranging from small-scale preps from one plate to concentrated virus stock produced on 9 plates.

2. Ready-to-transduce lentiviral preps - ready to use viral stocks of general utility, including lentiviral constructs for fluorescently labeling either whole cells or organelles, negative control vectors for shRNA constructs, and for cell reprogramming.

  • hPGK-eGFP
  • hPGK-mCherry
  • hPGK-dTomato
  • hPGK-Luciferase
  • Tet-inducible-eGFP
  • Tet-inducible-dTomato
  • CMV-NLS-eGFP (nuclear eGFP)
  • CMV-NLS-mCherry (nuclear mCherry)
  • CMV-eGFP-CAAX (membrane eGFP)
  • CMV-mCherry-CAAX (membrane mCherry)
  • CMV-mCherry-CAAX-2A-NLS-eGFP (membrane mCherry & nuclear eGFP)
  • LKO.1-scrambled shRNA (Sigma-Aldrich)
  • GIPZ-scrambled shRNA (Open Biosystems)
  • TRIPZ-scrambled shRNA (Open Biosystems)
  • iPSC 4-in-1 reprogramming lentivirus (Allele Biotechnology, 20μL/tube)

3. Full service lentiviral vector construction and viral particle production – Custom construction of a lentiviral vector for constitutive or inducible expression of either shRNA or cDNA, followed by viral packaging and titration, as well as ready-to-transduce control fluorescent lentivirus. Similar “the works” services are being initiated for retroviral and adenoviral vectors. Please contact the core to discuss potential full-service projects.

Other Core services

  • Custom viral particles concentration and purification with user-provided viral supernatants
  • Large scale DNA plasmid preparations
  • Ready-to-use polybrene (1 mg/mL, 100µL per aliquot, 5 aliquots)
  • Consultation

Equipment and Resources

The Core is fully equipped with all the necessary equipment for production of consistent quality viral vectors including state-of-the art cell culture hoods and incubators under appropriate biosafety conditions, as well as an ultracentrifuge, microscopes and a dedicated -80C freezer for storage of vectors. Producer cell lines 293T and 293A cells are routinely tested for the presence of mycoplasma using Lonza detection kit and discontinued at passage 20.

The Institute is in a partnership program with Sigma-Aldrich and Open Biosystems. Both companies provide affordable shRNA constructs. The Core also collaborates with Allele Biotechnology, to provide a ready-to-transduce 4-in-1 reprogramming lentivirus including iPSC. The Core uses the second or third generation protocol for VSVG pseudotyped lentiviral and retroviral production with standard 100x concentration and purification of virus by ultracentrifugation combined with sucrose cushion.

Contact

For questions and comments please contact:

Chun-Teng Huang, Core Manager
Room 7112
cthuang@sanfordburnham.org
(858) 646-3100 Ext. 4353

Price List
Biosafety level 2 virus shipping container$30.00$45.00$88.00
Concentrated virus packaging additional plateEach plate$100.00$135.00$263.00
Concentrated virus packaging, with qualitative titration3 plates$400$600.00$1,169
Concentrated virus packaging, with quantitative titration (using FACS)3 plates$525$788.00$1,535
Large scale DNA plasmid preparations$125.00$188.00$366.00
Ready-to-transduce lentiviral preps. (per vial)$55.00$83.00$161.00
Ready-to-use Polybrene (1 mg/mL, 100µL per aliquot, 5 aliquots)$10.00$15.00$30.00
Step1: Ligation/transformation discontinueStep 1$100.00$135.00$263.00
Step2-1: Miniprep evaluation discontinue (PCR confirmation)Step 1 and 2-1$150.00$203.00$395.00
Step2-2: Miniprep evaluation discontinue (sequencing)Step 1, 2-1, and 2-2$200.00$270.00$526.00
Step2-3: Miniprep evaluation discontinue (transfection)Step 1 and 2$250.00$338.00$658.00
Step3: Maxi-prep discontinueStep 1, 2, and 3$300.00$405.00$789.00
The works: Custom constitutively expressed cDNA lentiviral vectInsert size 1~2kb, 9 plates$1,100$1,485.00$2,893
The works: Custom constitutively expressed cDNA lentiviral vectorInsert size < 1kb, 9 plates$1,000.00$1,250.00$1,350.00$2,630.00
The Works: Custom constitutively expressed cDNA lentiviral vectorInsert size 2~3kb, 9 plates$1,200$1,800.00$3,507
The works: Custom constitutively expressed shRNA lentiviral vector1 shRNA, 9 plates$1,000$1,500.00$2,923
The works: Custom constitutively expressed shRNA lentiviral vector2 shRNA (targeting the same gene), 9 plates$1,800$2,430.00$4,734
The works: Custom constitutively expressed shRNA lentiviral vector3 shRNA (targeting the same gene), 9 plates$2,400$3,240.00$6,312
The works: Custom inducible cDNA lentiviral vectorInsert size < 1kb, 9 plates$1,000$1,350.00$2,630
The works: Custom inducible cDNA lentiviral vectorInsert size 1~2kb, 9 plates$1,100$1,485.00$2,893
The works: Custom inducible cDNA lentiviral vectorInsert size 2~3kb, 9 plates$1,200$1,620.00$3,156
Virus concentration and purification (per 135mL of unconcentrated soup$100$135.00$263
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