pLX-sgRNA
(Plasmid #50662)

• Purpose: Lentiviral expression of AAVS1-targeting sgRNA; insert can be replaced with custom sgRNA (see protocol)
• Depositing Labs: Eric Lander, David Sabatini
• Publication: Wang et al Science. 2014 Jan 3;343(6166):80-4. doi: 10.1126/science.1246981. Epub 2013 Dec 12.

Backbone

• Vector backbone: pLX304
• Backbone size w/o insert (bp): 7000
• Total vector size (bp): 7500
• Vector type: Mammalian Expression, Lentiviral, CRISPR
• Selectable markers: Blasticidin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: sgAAVS1
• Insert Size (bp): 450
• Promoter: U6

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XhoI (not destroyed)
• 3′ cloning site: NheI (not destroyed)
• 5′ sequencing primer: cgggtttattacagggacagcag
• 3′ sequencing primer: taccagtcaatctttcacaaattttgt

Resource Information

• Supplemental Documents:
  • Individual sgRNA cloning protocol
• A portion of this plasmid was derived from a plasmid made by: sgAAVS1 was cloned from Plasmid 41818: gRNA_AAVS1-T2
• Articles Citing this Plasmid:
  • 71 References

Terms and Licenses

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

Depositor Comments

For more information on OE-PCR please see: http://en.wikipedia.org/wiki/Overlap_extension_polymerase_chain_reaction)

gRNA target sequence GGGGCCACTAGGGACAGGAT