AAV-mIL-30 was generated by a helper free packaging system (AAV-DJ, Cell Biolabs, San Diego, CA, USA). mIL-30 cDNA was expressed and reversed from IFN-γ-treated mouse macrophages and cloned to the adeno-associated virus vector (pAAV-IRES-GFP). The cloned plasmid was co-transfected with pAAV-DJ and pHelper at a ratio of 1:1:1 into the packaging HEK293 cell line. Viral particles were purified and quantified as previously reported [8 (link),15 (link)]. AAV-mock is identical to the above but contains no transgene in the expression cassette.
Aav dj
Manufactured by Cell Biolabs
Sourced in United States
The AAV-DJ is a recombinant adeno-associated virus (AAV) vector that can be used for gene delivery applications. It is a versatile AAV serotype derived from multiple naturally occurring AAV serotypes. The AAV-DJ vector provides efficient transduction of a wide range of cell types and tissues.
Automatically generated - may contain errors
Lab products found in correlation
Phelper, by Cell Biolabs (2 mentions)
Hitrap heparin column, by GE Healthcare (1 mentions)
Gateway system, by Thermo Fisher Scientific (1 mentions)
Hitrap, by Cytiva (1 mentions)
Aavpro titration kit, by Takara Bio (1 mentions)
Aavpro purification kit, by Takara Bio (1 mentions)
Aavpro293 cells, by Takara Bio (1 mentions)
6 protocols using aav dj
1
Generation of AAV-mIL-30 Viral Particles
2
Production and Purification of Recombinant AAVs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
AAV was prepared as described previously (McClure et al., 2011 (link); Park et al., 2016 (link)). Briefly, three plasmids encoding the required components for AAV production were transfected into 293 AAV cells (Cell Biolab, Inc) using the calcium phosphate methods: AAV-DJ, pHelper (Cell biolabs, Inc), and pAAV-CaM kinase II. For production of MLI-specific AAVs (pX), a putative promoter region of pAAV-synapsin vector was replaced into promoter region of Nos-1, Grin3a, Kit and obtained pX-GFP and pX-cre AAVs using with Gateway system (Thermo Fisher Scientific). After 48–60 hr post-transfection, cells were solubilized and AAVs were purified using a HiTrap Heparin column (GE healthcare).
3
Recombinant AAV-DJ Expressing Murine IL-22
4
Astrocyte-Specific Gene Delivery Using AAV
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Vector XX680 was obtained from the University of Pennsylvania (Philadelphia, PA, USA). We replaced the existing promoter in pAAV-EF1a-DIO vector with an astrocyte-specific promoter (glial fibrillary acidic protein, GFAP). GFAP was derived from an Addgene plasmid hGFAP-NLS-HA-Dre-P2A-BFP [64 (link)]. We replaced the promoter after PCR amplification of the GFAP and digestion of it with MluI/BamHI restriction enzymes. mVenus was inserted between BamHI/EcoRI sites. Constructs were packaged into adeno-associated virus pseudotype DJ (AAV-DJ) (Cellbiolabs, San Diego, CA, USA) using the triple-transfection technique, as previously described [65 (link)]. Titers of each virus were set to be between 1 × 107–1 × 108 vector genomes/mL by qPCR using Takara AAVpro® Titration kit (Takara Bio Inc., Kusatsu, Shiga Prefecture, Japan).
5
Cloning and Packaging of AAV Vectors for Htr1a and BDNF Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cDNA encoding mouse Htr1a was cloned into the pAAV-Syn-eGFP vector. The packaging of DNA of the pAAV-Syn-Bdnf-eGFP plasmid or of the control pAAV-Syn-eGFP plasmid into rAAV capsids was performed using co-transfection with plasmids AAV-DJ and pHelper (Cell Biolabs, Inc., San Diego, CA, USA). Viral particles were harvested in 48 h according to the protocol described by Grimm and coauthors [96 (link)]. The amount of the obtained viral particles was determined with real-time quantitative PCR with primers F 5′-cctggttgctgtctctttatgagg and R 5′-tgacaggtggtggcaatgc. A series of dilutions of an original plasmid of known concentration was used as standards for determining the number of viral particles. All AAV vectors employed in this study had identical genomic titers (109 viral genomes per microliter).
6
AAV2-luc Transduction Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
AAV2-luc was purchased from Cell Biolabs (San Diego, CA). AAV vectors containing hEx45 sgRNAs #1 and #23 were constructed by cloning synthetic oligos (Eurofins Genomics, Tokyo, Japan) into pAAV-Guide-it-Down vector (Takara Bio, Shiga, Japan). pAAV-Guide-it Up vector was purchased from Takara Bio. These vectors were packaged into AAV-DJ (Cell Biolabs) using AAVpro293 cells (Takara Bio) and purified using an AAVpro Purification Kit (Takara Bio).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!