Aav dj packaging system

Manufactured by Cell Biolabs

Sourced in United States

The AAV-DJ Packaging System is a reagent kit designed for the production of recombinant adeno-associated virus (AAV) particles. The system provides the necessary components for efficient packaging of AAV vectors, including the AAV-DJ serotype, which is known for its ability to transduce a broad range of cell types. The kit includes plasmids and other required materials to facilitate the generation of high-titer AAV particles in a laboratory setting.

Automatically generated - may contain errors

Lab products found in correlation

Paav dj, by Cell Biolabs (1 mentions)

Spelling variants of this product

AAV-DJ Helper Free Packaging System (10 citations) AAV-DJ (6 citations)

3 protocols using aav dj packaging system

Recombinant AAV Production for B11-Fc Antibody

Cited 4 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

The AAV-DJ Packaging System (Cell Biolabs, USA) with the pAAV-DJ Vector, pHelper Vector, and pAAV-GFP Control Vector plasmids were used to construct plasmids for rAAV-B11-Fc production. The pAAV-DJ Vector contains the rep genes required for replication and the cap genes encoding capsid proteins. The pHelper Vector contains most of the adenoviral genome required for infectious rAAVs assembly (i.e., genes E2A, E4, and VA RNA).
The technique of Fc-fused single-domain antibody development was described previously (Godakova et al., 2019 (link); Esmagambetov et al., 2021 (link); Voronina et al., 2021 (link); Favorskaya et al., 2022 (link)). The nucleotide sequence encoding B11-Fc antibody was synthesized at Evrogen Company (Moscow, Russia) and cloned into the pAAV–EGFP Control Vector plasmid instead of the EGFP gene at the EcoRI and XbaI restriction sites, thus obtaining the pAAV-B11-Fc plasmid.

Derkaev A.A., Ryabova E.I., Esmagambetov I.B., Shcheblyakov D.V., Godakova S.A., Vinogradova I.D., Noskov A.N., Logunov D.Y., Naroditsky B.S, & Gintsburg A.L. (2022). rAAV expressing recombinant neutralizing antibody for the botulinum neurotoxin type A prophylaxis. Frontiers in Microbiology, 13, 960937.

+ Open protocol

+ Expand

Recombinant AAV Production in HEK293T

Check if the same lab product or an alternative is used in the 5 most similar protocols

HEK293T cells were seeded in 15 cm dishes and transfected at 80–90% confluence. GFP-AAV plasmid, N-ABE-AAV or C-ABE-AAV were transfected along with pHelper and pAAV-DJ from the AAV-DJ Packaging System from Cell Biolabs in a 1:1:1 ratio using calcium phosphate and a total of 60 µg per plate. Media was replaced 24 h post transfection. Cell pellets were harvested at 72 h post transfection through manual cell scraping and centrifuged at 1500×g for 12 min. After aspirating the supernatant, the cell pellet was resuspended in 1 mL AAV lysis buffer (50 mM Tris-HCl pH = 8.5, 150 mM NaCl and 2 mM MgCl₂). Resuspended pellets were subjected to three freeze–thaw cycles between an ethanol/dry ice bath and a 37 °C water bath. Lysed cell pellets were then spun at 10,000×g for 10 min and the supernatant was collected as crude lysate. Lysates were then treated with 50 U benzonase per mL and incubated at 37 °C for 30 min to digest unpackaged plasmid. Crude lysates were added directly to cells or flash frozen with liquid nitrogen and stored at −80 °C for future use.

Winter J., Luu A., Gapinske M., Manandhar S., Shirguppe S., Woods W.S., Song J.S, & Perez-Pinera P. (2019). Targeted exon skipping with AAV-mediated split adenine base editors. Cell Discovery, 5, 41.

+ Open protocol

+ Expand

Construction and Characterization of Viral Vectors

Check if the same lab product or an alternative is used in the 5 most similar protocols

The following mono-cistronic viral vectors with a synapsin promotor as well as bi-cistronic viral vectors with a H1 and a synapsin promotor were constructed: pAAV_Syn-eGFP, pAAV_Syn-eGFP-Tau[R406W], pAAV_H1-scramble-shRNA_Syn-eGFP, pAAV_H1-5-HT7R-shRNA_Syn-eGFP (Supplementary Fig. 8A,B,D). Following target sequences for the shRNA were used: scramble (scr) 5 ′ -ACTACCGTTGTTATAGGTG-3 ′ ; anti-5-HT7R 5 ′ -TCCTTTATCATTATGGTGT-3 ′ . Adeno-associated viruses (AAV) were produced using the AAV-DJ Packaging System from CELL BIOLABS in HEK293 cells. Briefly, HEK 293 cell were transfected with a pAAV plasmid encoding the gene of interest along with a pHelper plasmid as well as a pAAV-DJ plasmid. After three days, AAVs were harvested by three freezing and thawing cycles followed by benzonase digest. AAVs were concentrated using Amicon centrifugation tubes with a cut off of 10 kDa. For titer determination virus capsid was digested using protein K, and virus DNA was analyzed using quantitative realtime polymerase chain reaction (qRT-PCR) with primers against the viral WPRE element (fw 5 ′ -CCTGGTTGCTGTCTCTTTATGAGG-3 ′ ; rev 5 ′ -TGACAGGTGGTGGCAATGC-3 ′ ; probe 5 ′ -/6-FAM/ CGTTGTCAGG-CAACGTGG CGTGGTG/TAMRA/-3 ′ (Sigma). For calculation of the viral gene copy number per μl the relative standard curve method was used.

Labus J., Röhrs K.F., Ackmann J., Varbanov H., Müller F.E., Jia S., Jahreis K., Vollbrecht A.L., Butzlaff M., Schill Y., Guseva D., Böhm K., Kaushik R., Bijata M., Marin P., Chaumont-Dubel S., Zeug A., Dityatev A, & Ponimaskin E. (2021). Amelioration of Tau pathology and memory deficits by targeting 5-HT7 receptor. Progress in neurobiology, 197.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!