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, sindbis virus, AAV, and VSV 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 therapeutic target and drug screening. The viral core now integrates CRISPRs/Cas technology in a plasmid-free viral platform including lentivirus, adenovirus and AAV, providing scientists with choices of combined technologies to manipulate genes using CRISPR gene disruption, editing, inhibition, and activation. More importantly, the core actively participates and supports translational research to fight human diseases.
Custom packaged lenti-viral (HIV-1 and FIV) production is based on constructs provide by the customer. Titration is offered for virus ubiquitously expressing fluorescent marker.
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- 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)
"The Works" custom lentiviral vector construction – offers vector generation based on cDNA or shRNA of interests with a choice of constitutive/inducible expression vectors, viral production, and titration.
Custom viral particles concentration and purification with user-provided viral supernatants allows customers to increase viral titer (up to 1000 fold) based on crude viral supernatants.
Custom packaged retro-viral (MMLV) production is based on constructs provide by the customer. Titration is offered for virus ubiquitously expressing fluorescent marker.
Ready-to-transduce retroviral preps - include: constitutively expressed fluorescent protein.
• GFP
• mCherry
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) titration using the plaque assay.
"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.
"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
AAV-DJ : In vitro, liver, heart, kidney
AAV-DJ/8 : liver, heart, kidney, pancreas, SM, brain (i.v.)
• Ready-to-transduce AAV
AAV-2-GFP
AAV-DJ-GFP
All-in-one CRISPR/Cas gene disruption and editing viral vectors
• Knock-out using viral vector platform (hCas9 + gRNA)
Gene disruption using adenovirus or lentivirus (on demand)
Example: T7EI assay, gRNA-T1 to 3 targeting Rosa26 loci
Example: Ready-to-transduce Ad5-CRISPR all-in-1 targeting GFP
• 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
Example: knock-in GFP-2A-Gene of interests in Rosa26 loci
Genetic modified mouse creation
• Knock-in (hCas9 + gRNA + Donor DNA)
Rosa26 loci
Choice of C57BL/6 C2 and 129 mESC.
Example: knock-in GFP-2A-Gene of interests in Rosa26 loci
gRNA#1: 4 out of 4 screened mESC clonal line(100%) show Rosa loci integration
gRNA#2: 3 out of 4 screened mESC clonal line (75%) show Rosa loci integration
gRNA#3: 2 out of 4 screened mESC clonal line (50%) show Rosa loci integration
Custom CRISPR gene inhibition (CRISPRi) and activation (CRISPRa)
• CRISPRi viral vector delivery
Adenovirus (dCas9-Krab)
Lentivirus (dCas9-Krab and gRNA)
Example: SOX2 CRISPRi
• CRISPRa viral vector delivery
Lentivirus (dCas9-VP64 and gRNA)
Viral Vector Core at La Jolla has not received any reviews.
Viral Vector Core at La Jolla has not received any endorsements.