lentiCRISPRv2-sgRNAs-del-AK057321-SVA-mcherry
(Plasmid
#202821)
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PurposeLentiviral vector modified to express two sgRNAs that delete the SVA within the SVA-lncRNA AK057321 gene (exon 3) with EF-1apha for Cas9 and mcherry expression
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 202821 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbone98291
- Backbone size w/o insert (bp) 12817
- Total vector size (bp) 13590
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Modifications to backboneThis vector was modified from Addgene 98291. The hygromycin resistance gene was excised by digestion with PshAI and MluI and replaced with PCR-amplified mcherry from pLVX-mcherry. lentiCRISPRv2-mcherry was then digested with KpnI and NheI and PCR fragments containing two sgRNAs, each with their own U6 promoter were assembled using NEBuilder HiFi DNA Assembly (NEB).
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Vector typeMammalian Expression, Lentiviral
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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Copy numberUnknown
Gene/Insert
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Gene/Insert nametwo sgRNAs (each with a U6 promoter) to delete SVA within SVA-lncRNA AK057321 gene
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SpeciesH. sapiens (human)
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Insert Size (bp)773
- Promoter U6 (two)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site KpnI (not destroyed)
- 3′ cloning site NheI (not destroyed)
- 5′ sequencing primer whole plasmid sequencing
- 3′ sequencing primer whole plasmid sequencing (Common Sequencing Primers)
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:
lentiCRISPRv2-sgRNAs-del-AK057321-SVA-mcherry was a gift from Matthew Anderson (Addgene plasmid # 202821 ; http://n2t.net/addgene:202821 ; RRID:Addgene_202821) -
For your References section:
Hominoid SVA-lncRNA AK057321 targets human-specific SVA retrotransposons in SCN8A and CDK5RAP2 to initiate neuronal maturation. Nadler MJS, Chang W, Ozkaynak E, Huo Y, Nong Y, Boillot M, Johnson M, Moreno A, Matthew P Anderson. Commun Biol. 2023 Mar 30;6(1):347. doi: 10.1038/s42003-023-04683-8. 10.1038/s42003-023-04683-8 PubMed 36997626