TKIT NFL gRNA1 gRNA2
(Plasmid #182681)

• Purpose: Expression of spCas9, gRNA targeting intron 3 and gRNA targeting 3'UTR of the mouse Nefl gene. Can be used for the tagging of endogenous NFL via Targeted Knock-In with Two guides (TKIT) approach.
• Depositing Lab: Ivana Nikić-Spiegel
• Publication: Arsic et al Nat Commun. 2022 Jan 14;13(1):314. doi: 10.1038/s41467-022-27956-y.

Backbone

• Vector backbone: pORANGE
• Vector type: Mammalian Expression, CRISPR

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: two Nefl-targeting gRNAs, one targets intron 3 and the other 3'UTR of the Nefl gene
• Alt name: Nefl
• gRNA/shRNA sequence: ACATATCCTTTAGGAGAGTG and GTATAATTCTGAGATGACTT
• Species: M. musculus (mouse)
• GenBank ID:
• Entrez Gene: Nefl (a.k.a. CMT2E, NF-L, NF68, Nfl)
• Promoter: U6 and chicken beta-actin promoter

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: NA
• 3′ sequencing primer: NA

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Following pORANGE cloning template vector (https://www.addgene.org/131471/) was used as a vector backbone.

Terms and Licenses

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