Viral Vectors Lab Equipment

Viral Vectors

Assisting Institute investigators with cost-effective production of high-quality, high-titer lentiviral, retroviral and adenoviral vector.

 

The viral vector core facility at Sanford Burnham Prebys Medical Discovery Institute develops state-of-the-art viral vector-based gene delivery technology. Its portfolio ranges from lentivirus, retrovirus, adenovirus, AAV, VSV, sindbis virus, and Zika virus products as well as customized “The Works” viral vector construction and swapping service package. There are a variety of ready-to- transduce viral vector collections including constitutively expressed or inducible fluorescent proteins for nuclear or membrane localization and bioluminescence reporters for 3D drug screening. The viral core has integrated CRISPRs/Cas technology in the plasmid-free viral vector platform, providing scientists with choices of combined technologies to manipulate genes using CRISPR gene disruption, editing, inhibition, and activation. In addition, we extend technical expertise in nanoparticle research including exosome purification, analysis, and engineering. More importantly, the core actively participates and supports translational research to fight human diseases.

 

The Viral vector core facility focuses on the following services:
 

  1. Custom exosome purification, analysis, and engineering
    • Exosome isolation from user-provided cell culture supernatant or biological fluids.  Options of large scale exosome concentration and purification include (1) differential untracentrifugation (2) density gradient (3) precipitation* (4) immunoaffinity* (on demand).
    • SBI XPACK exosome loading technology*
    • SBI exosome tracking technology*
    • SBI exosome miRNA array analysis*
         *SBP viral vector core facility is in partnership with System Biosciences Inc. (SBI).
  2. Custom CRISPR gene inhibition (CRISPRi) and activation (CRISPRa)
    • Lentiviral vector delivery (MTA)
  3. All-in-one CRISPR/Cas gene disruption and editing viral vectors
    • Knock-out using viral vector platform (hCas9 + gRNA)
    • Knock-in using plasmid-free viral vector platform (hCas9 + gRNA + Donor DNA)
      • Gene editing using adenovirus (on demand)
      • Human AAVS1 safe harbor loci
      • Mouse Rosa26 safe harbor loci
  4. Genetic modified mouse creation
    • Knock-in (hCas9 + gRNA + Donor DNA)
      • Rosa26 loci
      • Choice of C57BL/6 C2 and 129 mESC
  5. "The Works" custom AAV vector construction and virus packaging
    • Serotype Tropism
      • AAV-1 :  SM, CNS, retina, pancreas
      • AAV-2 :  In vitro, VSMC, SM, CNS, liver, kidney
      • AAV-3 :  Hepatocarcinoma, SM
      • AAV-4 :  CNS, retina, lung
      • AAV-5 :  SM, CNS, lung, retina
      • AAV-6 :  SM, SM (i.v.), heart, lung
      • AAV-8 :  Liver, SM, CNS, retina, pancreas, heart (MTA and collaboration)
      • AAV-DJ : In vitro, liver, heart, kidney (MTA and collaboration)
      • AAV-DJ/8 : liver, heart, kidney, pancreas, SM, brain (i.v.) (MTA and collaboration)
      • AAV-PHP.B, AAV-B1, and Retro-AAV (MTA and collaboration)
    • Ready-to-transduce AAV
      • AAV-2-GFP
      • AAV-8-GFP (MTA)
      • AAV-DJ-GFP (MTA)
      • AAV- DJ/8-GFP (MTA)
      • AAV- PHP.B-GFP (MTA)
  6. Custom packaged lentiviral (HIV-1 and FIV) and retroviral production is based on constructs provide by the customer. Titration is offered for virus ubiquitously expressing fluorescent marker.
     
  7. Custom packaged integrase defective lenti-virus (IDLV) production
     
  8. Custom production of adenoviral particles (Ad5) from user-supplied construct or virus - includes: (1) adenovirus recovery (2) high purity adenoviral preparation using CsCl gradient purification (3) Additional adenoviral column concentration and buffer exchange (4) titration using the plaque assay.
     
  9. "The Works" custom lentiviral-adenoviral vector construction and swapping - includes: (1) constitutive expression of custom cDNAs (2) inducible shRNA of interests expressed under a new Tet inducible system (3) Custom promoters/enhancer reporter vectors (4) MicroRNA sponges.
     
  10. Ready-to-transduce lentiviral preps - include: (1) constitutively expressed or inducible fluorescent protein, antibiotic selection marker, or bioluminescence reporter (2) nuclear or membrane localization tool (3) constitutively expressed non-silencing shRNA control.
    • hPGK-eGFP
    • hPGK-mCherry
    • hPGK-H2B-eGFP (nuclear eGFP)
    • hPGK-H2B-mCherry (nuclear mCherry)
    • hPGK-tdTomato
    • hPGK-Firefly Luciferase
    • hEF1α-tdTomato-T2A-Firefly Luciferase
    • UBC-eGFP-Firefly Luciferase fusion
    • CMV-Renilla-Luciferase
    • Tet-inducible-eGFP
    • Tet-inducible-tdTomato
    • CMV-NLS-eGFP (nuclear eGFP)
    • CMV-NLS-mCherry (nuclear mCherry)
    • CMV-eGFP-CAAX (membrane eGFP)
    • CMV-mCherry-CAAX (membrane mCherry)
    • LKO.1-scrambled shRNA (Sigma-Aldrich)
    • GIPZ-scrambled shRNA (Open Biosystems)
    • TRIPZ-scrambled shRNA (Open Biosystems)
  11. Custom lenti- and adeno-viral reporter for 2D and 3D screening
    • One secreted luciferase is constitutively expressed by hPGK promoter for normalization.
    • Another secreted luciferase is expressed by your promoter of interest.
    • The third secreted marker is expressed by another promoter of interest, your choice.
  12. Cell Cycle Reporter
    • Lenti and AAV viral vector delivery (MTA)
  13. Custom Zika virus production and titration
    • Zika strain (on demand)
      • African
      • Asian
      • Brazilian (2015-2016 outbreak)
      • Panama
    • Requirements
      • IBC approval
      • Collaboration and MTA
      • Safety training, work at BSL2+
  14. "The Works" custom VSV vector construction and production
    • Pseudotyped VSV-ΔG-GFP
    • Ebola virus glycoprotein (EboV-GP) Zaire strain (Kissidougou-C15, 2014)
    • Vesicular stomatitis virus glycoprotein (VSV-G)
  15. Custom packaged Sindbis viral vector production
     
  16. Ready-to-use zbFGF (zebrafish basic FGF) is a substitute for human bFGF on hES cell culture. High quality and quantity with less cost. Quality control is done batch by batch before releasing and the dilution of zbFGF is tested on hES cells.
     
  17. Other core services
    • Custom viral particles concentration and purification with user-provided viral supernatant allows customers to increase viral titer (up to 1000X) based on crude viral supernatant.
    • 100X viral concentration kit provides the concentration of crude viral lysates from small-scale viral production. Our kit is quick, easy, and allows for the long term storage of your virus at -80ºC.
    • Large scale DNA plasmid preparation (ready for viral production) starts from a small amount of customer's vector (e.g. miniprep) or transformed bacterial stock.
    • Ready-to-use polybrene - (1 mg/mL, 100µL per aliquot, 5 aliquots)
    • Consultation is free for core users.

 

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 -80°C 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 currently collaborates with Dr. Terskikh and Dr. Mercola labs to increase ready-to-transduce virus inventory.

 

For a Price List, please call (858)646-3100 ext. 4353 or email us.

Leadership

Associate Director

Contact

Chun-Teng Huang
Room 7112
(858) 646-3100 Ext. 4353
 

Vivian Lin
Room 7112
(858) 646-3100 Ext. 3110


 

Contact the Facility

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