pDGE Dicot Genome Editing Kit
(Kit #
1000000084
)
Depositing Lab: Johannes Stuttmann
Golden Gate cloning-based assembly of constructs for CRISPR genome editing in dicotyledonous plants.
Please Note: Plasmids pDGE144-169 are not functional for genome editing in Arabidopsis thaliana. The Stuttmann lab has more recent vectors providing improved efficiencies and/or selection markers, which may be obtained by directly contacting Johannes Stuttmann ([email protected]).
This kit will be sent as bacterial glycerol stocks in 96-well plate format.
Original Publication
Generation of chromosomal deletions in dicotyledonous plants employing a user-friendly genome editing toolkit. Ordon J, Gantner J, Kemna J, Schwalgun L, Reschke M, Streubel J, Boch J, Stuttmann J. Plant J. 2016 Aug 31. doi: 10.1111/tpj.13319. [Epub ahead of print] PubMed PMID: 27579989.
Description
This collection was designed for the simple Golden Gate cloning-based assembly of constructs for genome editing in dicotyledonous plants. The collection consists of “genome editing recipient” and “sgRNA shuttle” vectors, respectively. Recipient vectors are Agrobacterium-compatible, and contain a Cas9 expression cassette, a plant-selectable marker and a /ccdB/ cassette as a “dummy”. Shuttle vectors allow the generation of sgRNA transcriptional units from annealed oligonucleotides. sgRNA transcriptional units are then mobilized into recipient vectors, replacing the /ccdB/ dummy.
The Modular Cloning Toolkit (Addgene #1000000044), Modular Cloning Plant Parts (Addgene #1000000047) and hCas9 modules (Addgene #49771 and #49770) provided by Sylvestre Marillonnet, Nicola Patron and Sophien Kamoun, respectively, were used in construction of pDGE/pJOG plasmids.
Depositor-provided Files
The Stuttmann Lab has provided a manual that includes information about:
- Detailed vector descriptions and architecture
- Selection and design for gRNAs
- Cloning protocol
The following publication features functional data using plasmids featured in this kit:
Dissection of TALE-dependent gene activation reveals that they induce transcription cooperatively and in both orientations. Streubel J, Baum H, Grau J, Stuttmann J, Boch J. PLoS One. 2017 Mar 16. doi: 10.1371/journal.pone.0173580. PubMed PMID: 28301511.
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 pDGE Dicot Genome Editing Kit was a gift from Johannes Stuttmann (Addgene kit # 1000000084).”
For your Reference section:
Generation of chromosomal deletions in dicotyledonous plants employing a user-friendly genome editing toolkit. Ordon J, Gantner J, Kemna J, Schwalgun L, Reschke M, Streubel J, Boch J, Stuttmann J. Plant J. 2016 Aug 31. doi: 10.1111/tpj.13319. [Epub ahead of print] PubMed PMID: 27579989.
pDGE Dicot Genome Editing Kit - #1000000084
- Resistance Color Key
Each circle corresponds to a specific antibiotic resistance in the kit plate map wells.
- Inventory
Searchable and sortable table of all plasmids in kit. The Well column lists the plasmid well location in its plate. The Plasmid column links to a plasmid's individual web page.
- Kit Plate Map
96-well plate map for plasmid layout. Hovering over a well reveals the plasmid name, while clicking on a well opens the plasmid page.
Resistance Color Key
| Chloramphenicol and Ampicillin | |
| Chloramphenicol and Spectinomycin | |
| Spectinomycin |
Inventory
| Well | Plasmid | Resistance |
|---|---|---|
| A / 1 | pDGE5 | Chloramphenicol and Ampicillin |
| A / 2 | pDGE7 | Chloramphenicol and Ampicillin |
| A / 3 | pDGE8 | Chloramphenicol and Ampicillin |
| A / 4 | pDGE9 | Chloramphenicol and Ampicillin |
| A / 5 | pDGE10 | Chloramphenicol and Ampicillin |
| A / 6 | pDGE11 | Chloramphenicol and Ampicillin |
| A / 7 | pDGE12 | Chloramphenicol and Ampicillin |
| A / 8 | pDGE13 | Chloramphenicol and Ampicillin |
| A / 9 | pDGE14 | Chloramphenicol and Ampicillin |
| A / 10 | pDGE15 | Chloramphenicol and Ampicillin |
| A / 11 | pDGE6 | Chloramphenicol and Ampicillin |
| A / 12 | pJOG294 | Chloramphenicol and Ampicillin |
| B / 1 | pDGE1 | Chloramphenicol and Spectinomycin |
| B / 2 | pDGE2 | Chloramphenicol and Spectinomycin |
| B / 3 | pDGE3 | Chloramphenicol and Spectinomycin |
| B / 4 | pDGE4 | Chloramphenicol and Spectinomycin |
| B / 5 | pDGE62 | Chloramphenicol and Spectinomycin |
| B / 6 | pDGE63 | Chloramphenicol and Spectinomycin |
| B / 7 | pDGE64 | Chloramphenicol and Spectinomycin |
| B / 8 | pDGE65 | Chloramphenicol and Spectinomycin |
| B / 9 | pDGE144 | Chloramphenicol and Spectinomycin |
| B / 10 | pDGE146 | Chloramphenicol and Spectinomycin |
| B / 11 | pDGE148 | Chloramphenicol and Spectinomycin |
| B / 12 | pDGE151 | Chloramphenicol and Spectinomycin |
| C / 1 | pDGE160 | Chloramphenicol and Spectinomycin |
| C / 2 | pDGE161 | Chloramphenicol and Spectinomycin |
| C / 3 | pDGE164 | Chloramphenicol and Spectinomycin |
| C / 4 | pDGE165 | Chloramphenicol and Spectinomycin |
| C / 5 | pDGE166 | Chloramphenicol and Spectinomycin |
| C / 6 | pDGE167 | Chloramphenicol and Spectinomycin |
| C / 7 | pDGE168 | Chloramphenicol and Spectinomycin |
| C / 8 | pDGE169 | Chloramphenicol and Spectinomycin |
| C / 9 | pDGE76 | Chloramphenicol and Spectinomycin |
| C / 10 | pDGE77 | Chloramphenicol and Spectinomycin |
| C / 11 | pDGE78 | Chloramphenicol and Spectinomycin |
| C / 12 | pDGE79 | Chloramphenicol and Spectinomycin |
| D / 1 | pJOG250 | Chloramphenicol and Spectinomycin |
| D / 2 | pJOG251 | Chloramphenicol and Spectinomycin |
| D / 3 | pJOG252 | Chloramphenicol and Spectinomycin |
| D / 4 | pJOG253 | Chloramphenicol and Spectinomycin |
| D / 5 | pJOG292 | Spectinomycin |