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
- Depositing Labs
- Publication
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | ||
---|---|---|---|---|---|---|
Pooled Library | 79153 | pC004 - CRISPR Cas13a PFS Library † | 1 | $275 | Add to Cart |
-
† A Cas13a/C2c2 plasmid is not included with this item. Can be used in conjuction with pC002 - LShC2C2 (Addgene #79153) or similar.
Library Details
-
Protospacer Flanking Sites4096
Library Shipping
The library is delivered in a microcentrifuge tube on blue ice. The tube's contents will not necessarily be frozen. For best results, minimize freeze-thaws.
-
Volume∼10 µL
-
Concentration10 ng/µL
Resource Information
-
Depositor Data
- GenBank File (2 MB) - The protospacer is represented as NNNNNN.
-
Terms and Licenses
Depositor Comments
The plasmids in this library each represent a target site for Cas13a that has 6 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 4096 (46) combinations of 6 nucleotides.
These pooled libraries were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the pooled libraries were described, and include Addgene in the Materials and Methods of your future publications.
Example for your Materials & Methods section:
pC004 - CRISPR PFS Cas13a library was a gift from Feng Zhang (Addgene # 79153)For your References section:
C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector. Abudayyeh OO, Gootenberg JS, Konermann S, Joung J, Slaymaker IM, Cox DB, Shmakov S, Makarova KS, Semenova E, Minakhin L, Severinov K, Regev A, Lander ES, Koonin EV, Zhang F Science. 2016 Jun 2. pii: aaf5573. PubMed 27256883