pC004 - CRISPR Cas13a/C2c2 Protospacer Flanking Site (PFS) Library
(Pooled Library #79153)

• Purpose:

  A library of protospacer flanking sites (PFS) inserted at the 5' end of the β-lactamase gene for screening PFS preference for Cas13a/C2c2.

• Vector Backbone:

  pUC19 (Plasmid #50005)

• Depositing Labs: Feng Zhang
• Publication: Abudayyeh et al Science. 2016 Jun 2. pii: aaf5573.

Library Details

• Protospacer Flanking Sites: 4096

Library Shipping

Each library is delivered in microcentrifuge tubes on blue ice. A tube's contents will not necessarily be frozen. For best results, minimize freeze/thaw cycles.

• Volume: ~10 µL
• Concentration: 10 ng/µL

Resource Information

• Protocols:
  • For the most up-to-date and detailed instructions on using this library, please see Abudayyeh, et al. 2016 .
• Depositor Data:
  • Library Vector (GB, 6.2 KB) - The protospacer is represented as NNNNNN.

Terms and Licenses

Academic/Nonprofit Terms:

• UBMTA

Industry Terms:

• Not Available to Industry

Trademarks:

• Zeocin® is an InvivoGen trademark.

Depositor Comments

The plasmids in this library each represent a target site for Cas13a that has six randomized bases on the 3' end to survey for protospacer flanking site (PFS) motifs that promote binding and cutting activity by Cas13a. The target site with random bases is inserted at the 5' end of the beta lactamase transcript so that when the transcript is cleaved by Cas13a, resistance to ampicillin is lost and the bacteria that are transformed with these constructs will die. The plasmids in the surviving population can then be sequenced and the PFS motifs that promote cleavage can be inferred from the depleted sequences.

The protospacers contained in the library represent all 4,096 (4⁶) combinations of six nucleotides.