AdenoBuilder toolkit
(Kit #
1000000176
)
Depositing Lab: Fred Bunz
Plasmids containing modular inserts that span the human adenovirus serotype 5 (HAdV5) genome that can be assembled in a one-tube reaction.
This kit will be sent as individual bacterial stabs.
Original Publications
Seamless assembly of recombinant adenoviral genomes from high-copy plasmids. Miciak JJ, Hirshberg J, Bunz F. PLoS One. 2018 Jun 27;13(6):e0199563. doi: 10.1371/journal.pone.0199563. PubMed PMID: 29949649
AdenoBuilder: A platform for the modular assembly of recombinant adenoviruses. Jang Y, Bunz F. STAR Protoc. 2022 Jan 20;3(1):101123. doi: 10.1016/j.xpro.2022.101123. PubMed PMID: 35098167
Description
These plasmids contain modular inserts that span the genome of human adenovirus serotype 5 (HAdV5). In a one-tube reaction, plasmid inserts are seamlessly assembled into full-length adenovirus genomes, which can then be directly packaged into infectious particles and amplified. The compact size of the individual modules facilitates the rapid and efficient modification of virtually any viral element. A unique attribute of this system is that mutations/alterations/transgene insertion in multiple viral segments can be combined into a single genome.
Figure 1. Segments of the Ad5 genome were cloned into high-copy plasmids. Each module, or “block”, contains overlapping sequences at either end (represented by distinct colors) that permit enzymatic fusion with the adjacent block. Assembled genomes are then delivered to standard packaging cells for preparation of a primary virus stock.
Figure 2. The plasmid inserts (“blocks”) span the adenoviral genome.
Figure 3. Modified blocks facilitate the insertion of transgenes into recombinant adenoviral vectors. In principle, transgenes up to 8.5 kb can be inserted into the E1 region. Deletions in the E3 and E4 regions can be incorporated to the final assembled genome so that the total size does not exceed the packaging capacity.
Protocol
AdenoBuilder: A platform for the modular assembly of recombinant adenoviruses. Jang Y, Bunz F. STAR Protoc. 2022 Jan 20;3(1):101123. doi: 10.1016/j.xpro.2022.101123. PubMed PMID: 35098167
How to Cite this Kit
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which they were created, and include Addgene in the Materials and Methods of your future publications.
For your Materials and Methods section:
“The AdenoBuilder toolkit was a gift from Fred Bunz (Addgene kit #1000000176).”
For your Reference section:
Seamless assembly of recombinant adenoviral genomes from high-copy plasmids. Miciak JJ, Hirshberg J, Bunz F. PLoS One. 2018 Jun 27;13(6):e0199563. doi: 10.1371/journal.pone.0199563. PubMed PMID: 29949649
AdenoBuilder: A platform for the modular assembly of recombinant adenoviruses. Jang Y, Bunz F. STAR Protoc. 2022 Jan 20;3(1):101123. doi: 10.1016/j.xpro.2022.101123. PubMed PMID: 35098167
The AdenoBuilder toolkit contains 13 plasmids. Please refer to the individual plasmid pages below for more details on each plasmid in this kit: