Viral Vector Core at La Jolla

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)