Cited by 572Virus-based gene therapy researchCRISPR and Genetic EngineeringRNA Interference and Gene Delivery

A public genome-scale lentiviral expression library of human ORFs

Open access
Yang, Xiaoping, Boehm, Jesse S, Yang, Xinping, Salehi-Ashtiani, Kourosh, Hao, Tong, Shen, Yun, Lubonja, Rakela, Thomas, Sapana R +16 more·2011·Nature Methods

Abstract

Functional characterization of the human genome requires tools for systematically modulating gene expression in both loss-of-function and gain-of-function experiments. We describe the production of a sequence-confirmed, clonal collection of over 16,100 human open-reading frames (ORFs) encoded in a versatile Gateway vector system. Using this ORFeome resource, we created a genome-scale expression collection in a lentiviral vector, thereby enabling both targeted experiments and high-throughput screens in diverse cell types.

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  • Current CRISPR sgRNA libraries can be decoded directly via the CROP-seq vector, while other perturbations, such as ORFs or non-coding elements, can be tagged with short barcodes.

    “Existing CRISPR sgRNA libraries can be read out directly (using the CROP-seq vector) while other perturbations (e.g., ORFs, non-coding sequences) can be paired with short barcodes (Melnikov et al., 2012; Yang et al., 2011).”

    From Optical Pooled Screens in Human Cells (2019) · cites this paper as Yang et al., 2011

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  1. [3]Moody SE Boehm JS Barbie DA Hahn WC Cur Opin Mol Ther 12 284 293 2010 20521217

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Authors
Yang, X., Boehm, J. S., Yang, X., Salehi-Ashtiani, K., Hao, T., Shen, Y., Lubonja, R., Thomas, S. R., Alkan, O., Bhimdi, T., Green, T. M., Johannessen, C. M., Silver, S. J., Nguyen, C., Murray, R. R., Hieronymus, H., Balcha, D., Fan, C., Lin, C., Ghamsari, L., Vidal, M., Hahn, W. C., Hill, D. E., & Root, D. E.
Affiliations
  • Broad Institute (US)
  • Harvard University (US)
Corresponding author
  • Vidal, Marc
  • Hahn, William C
  • Hill, David E
  • Root, David E
Venue
Nature Methods
Published
2011-06-26
Volume
8
Issue
8
Pages
659-661
ISSN
  • 1548-7091 (ISSN-L)
  • 1548-7105
OpenAlex
W2023994677
Cited by
570
References
20
Citations by year
2026: 35 · 2025: 40 · 2024: 35 · 2023: 49 · 2022: 26 · 2021: 48 · 2020: 52 · 2019: 58 · 2018: 40 · 2017: 35 · 2016: 40 · 2015: 44 · 2014: 31 · 2013: 18 · 2012: 15
FWCI
13.57
Type
journal article
Language
en
Open access
Locations
  • Nature Methods (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
Funders
National Cancer Institute (P01CA142536, R33CA128625)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
Keywords
ORFeome, Gateway cloning, loss-of-function screen, gain-of-function screen, functional genomics, mammalian cell expression
MeSH terms
Cloning, Molecular, Genetic Vectors, Humans, Genomic Library, Lentivirus, Open Reading Frames

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