Mouse CRISPR Activation Pooled Library (Caprano P65-HSF)
(Pooled Library #1000000113, #92383, #92384)
This mouse CRISPR activation library (Caprano P65-HSF) is in backbone pXPR_502 (P65-HSF). This Caprano library activates over 22,000 mouse genes and is used for genome-wide activation screening.
Each gene activated by this library is targeted by 3-6 gRNAs. These gRNAs are split among the two pools: Set A (Cat# 92383) contains gRNAs 1-3, Set B (Cat# 92384) contains gRNAs 4-6. Each pool has a unique set of gRNAs, but both pools are designed to target the same genes. The slight discrepancy in number of genes targeted between Set A and Set B is because not all genes had 6 gRNAs.
Depositor comments: The modified sgRNA tracr has two MS2 loops and two PP7 loops, although only the PP7 loops are used in this library. This library uses two vectors instead of 3, as the PP7-P65-HSF is on same vector as sgRNA itself. It was found that MS2-P65-HST (hygroR) vector gave poor titer. The 6 guides per gene were designed first based on location relative to TSS, then sequence preferences.
Primers for the lentiGuide backbone can be used with this library in the Sequencing Protocol.
pXPR_502, which uses P65-HSF to activate a gRNA-scaffold.
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:Mouse Caprano CRISPR activation pooled library set A was a gift from David Root and John Doench (Addgene #92383).
Mouse Caprano CRISPR activation pooled library set B was a gift from David Root and John Doench (Addgene #92384).
For your References section:Up, down, and out: next generation libraries for genome-wide CRISPRa, CRISPRi, and CRISPR-Cas9 knockout genetic screens. Sanson KR, Hanna RE, Hegde M, Donovan KF, Strand C, Sullender ME, Vaimberg EW, Goodale A, Root DE, Piccioni F, Doench JG. bioRxiv 356626. doi: 10.1101/356626. bioRxiv 356626