Tol2-LyzC-Rac-DN(guide resistant)-2a-mcherry-U6ac-Rac guides
(Plasmid
#168243)
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Purpose"Dominant negative control of the assay to rescue Rac expression in neutrophil specific knockout"
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 168243 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepME-GFP
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Modifications to backboneremove GFP and add Rac2(guide resistant)-2a-mcherry
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Vector typeZebrafish expression
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Copy numberUnknown
Gene/Insert
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Gene/Insert nameRac(guide resistant)-DN
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gRNA/shRNA sequencegggccaatgcgagaccctgt/gggctgtatcccagagtccc
- Promoter LyzC
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Tag
/ Fusion Protein
- mcherry (N terminal on insert)
Cloning Information
- Cloning method Gateway Cloning
- 5′ sequencing primer CGGGCCCCCCCTCGAGGCCACCATGGTGAGCAAGG
- 3′ sequencing primer CTCCACCGCGGCCGCTTAGAGCATCACGCAGCCC (Common Sequencing Primers)
Resource Information
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Supplemental Documents
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
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For your Materials & Methods section:
Tol2-LyzC-Rac-DN(guide resistant)-2a-mcherry-U6ac-Rac guides was a gift from Qing Deng (Addgene plasmid # 168243 ; http://n2t.net/addgene:168243 ; RRID:Addgene_168243) -
For your References section:
A robust and flexible CRISPR/Cas9-based system for neutrophil-specific gene inactivation in zebrafish. Wang Y, Hsu AY, Walton EM, Park SJ, Syahirah R, Wang T, Zhou W, Ding C, Lemke AP, Zhang G, Tobin DM, Deng Q. J Cell Sci. 2021 Apr 15;134(8). pii: 237799. doi: 10.1242/jcs.258574. Epub 2021 Apr 22. 10.1242/jcs.258574 PubMed 33722979