Zhang Lab - Multiplexed Overexpression of Regulatory Factors (MORF) Library
(Pooled Library #1000000218, #192821)

• Purpose:

  The Multiplexed Overexpression of Regulatory Factors (MORF) pooled library is a comprehensive collection of human transcription factor open reading frames. Each isoform is uniquely barcoded with a 24 bp sequence, to simplify factor identification during screening. Individual plasmids are also available on the MORF Collection page.

• Vector Backbone:

  pLX_TRC317 - Constructed by inserting ORF and barcode in pLEX_307

• Depositing Labs: Feng Zhang
• Publication: Zhang et al. Cell. 2023 Jan 5; 186(1):209-229. doi: 10.1016/j.cell.2022.11.026.

Library Details

• Species: Human
• ORFs: 3,548 isoforms of 1,836 genes
• Insert size range: 250 - 10,000 bp
• Lentiviral Generation: 3rd

Library Shipping

This library is delivered as suspended DNA in a microcentrifuge tube on blue ice. The tube's contents will not necessarily be frozen. For best results, minimize freeze-thaws.

For the Kit with GFP and mCherry controls ONLY: You will also receive bacterial stabs of TFORF3549 (GFP, Addgene ID #145025) and TFORF3550 (mCherry, Addgene ID #145026) plasmids. Please do NOT order these plasmids in addition to this library.

• Volume: ∼20 µl
• Concentration: 50 ng/µL

Resource Information

• Protocols:
  • TF ORF barcode PCR for NGS QC and bulk screening (85 KB)
  • TF ORF barcode dial out PCR for 10X scRNA-seq (59.2 KB)
• Depositor Data:
  • NGS Primer Table (11 KB)
  • Table of ORF accessions and sequences (13.4 MB)
  • With GFP/Cherry NGS Read Counts (104 KB)
  • Without GFP/Cherry NGS Read Counts (106 KB)
  • pLX_TRC317 Genbank File (15 KB)
• Terms and Licenses:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
  • Institut Pasteur Label License for cPPT
  Industry Terms:
  • Not Available to Industry

Depositor Comments

The Pooled library #137000 contains low levels of the TFORF3549 (GFP) and TFORF3550 (mCherry) control plasmids since cells that express the control plasmids will grow faster than cells with differentiated phenotypes in a pooled screen. The individual GFP and mCherry plasmids are also provided as bacterial agar stabs in the kit. If you have difficulty amplifying the control plasmids or if you would like higher amounts of mCherry or GFP, you can choose to add in more control plasmids separately, after library amplification. For example, add 3 ng of each control plasmid (GFP and mCherry) to 10 ug of the amplified pooled library.

For library transformation and amplification, please refer to Joung et al. Nat Protoc. 2017, with 2 modifications:

• 1. As this is an ORF library, increasing the coverage (i.e., colonies/ORF) to 5-10 electroporations is recommended to minimize skewing.
• 2. Plate 1 electroporation onto 2 large LB agar plates to help minimize colony contact.

• Multiplexed Overexpression of Regulatory Factors (MORF) Library