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PurposeExpression of CRISPRi with U6-gRNA (Bsa1 sites)
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 163024 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backboneAAV2
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Backbone manufacturerCell Biolabs
- Backbone size w/o insert (bp) 3500
- Total vector size (bp) 7188
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Vector typeMammalian Expression, AAV
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)NEB Stable
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Growth instructions34-35 degrees is ideal
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameMammalian codon-optimized SadCas9
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Alt namenuclease deactivated SauCas9
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SpeciesS. aureus
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Insert Size (bp)3681
- Promoter hEF-1a
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Tags
/ Fusion Proteins
- KRAB (N terminal on insert)
- myc NLS (C terminal on insert)
- SV40 polyA (C terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site EcoR1 (not destroyed)
- 3′ cloning site Kpn1 (not destroyed)
- 5′ sequencing primer Mlu-EF1-Seq-S
- 3′ sequencing primer Add-Avr2-A (Common Sequencing Primers)
Resource Information
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Supplemental Documents
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A portion of this plasmid was derived from a plasmid made byKeith Joung (S. aureus dCas9 from Addgene #70703).
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:
(559) pAAV EF-1a KRAB-SadCas9 U6-gRNA was a gift from Tonia Rex (Addgene plasmid # 163024 ; http://n2t.net/addgene:163024 ; RRID:Addgene_163024) -
For your References section:
Optimization of S. aureus dCas9 and CRISPRi Elements for a Single Adeno-Associated Virus that Targets an Endogenous Gene. Backstrom JR, Sheng J, Wang MC, Bernardo-Colon A, Rex TS. Mol Ther Methods Clin Dev. 2020 Sep 6;19:139-148. doi: 10.1016/j.omtm.2020.09.001. eCollection 2020 Dec 11. 10.1016/j.omtm.2020.09.001 PubMed 33024795