AAV-U6/TO-shEsr1-CMV-TetR-eGFP
(Plasmid #233298)

• Purpose: DOX-inducible U6 driven shRNA against mouse Esr1
• Depositing Lab: Dayu Lin
• Publication: Golden et al bioRxiv [Preprint]. 2024 Sep 25:2023.12.09.570945. doi: 10.1101/2023.12.09.570945.

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Backbone

• Vector backbone: pJEP10-AAV-U6/TO-gRNA(Empty)-CMV-TetR-P2A-eGFP-KASH-pA
• Backbone manufacturer: Jonathan Ploski (Addgene #82706)
• Backbone size w/o insert (bp): 6298
• Total vector size (bp): 6240
• Modifications to backbone: Remove the sequence of gRNA scaffold
• Vector type: Mammalian Expression, AAV

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Esr1 shRNA
• Alt name: ERα
• gRNA/shRNA sequence: GGC ATG GAG CAT CTC TAC A
• Species: M. musculus (mouse)
• Entrez Gene: Esr1 (a.k.a. ER, ER-alpha, ERa, ERalpha, ESR, Estr, Estra, Nr3a1)
• Promoter: U6

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SapI (not destroyed)
• 3′ cloning site: XbaI (not destroyed)
• 5′ sequencing primer: hU6-F

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

There is a D insertion after T52 upstream of TetR and does not affect function. The expression of TetR was confirmed using Western Blotting in in vitro cell culture experiment.