Nature CommunicationsSystems BiologyPeer-reviewed journal article
Multiplexed pooled library screening with Cpf1
Jintan LiuiD, Sanjana SrinivasaniD, Chieh-Yuan LiiD, I-Lin HoiD, Johnathon Lynn RoseiD, MennatAllah ShaheeniD, Gang WangiD, Wantong YaoiD, Angela K. DeemiD, Chris Bristow, Traver HartiD, Giulio Francesco DraettaiD
Abstract
RNA interference and CRISPR/Cas9-based pooled library screens have revolutionized the field of functional genomics. However, currently available pooled library screens face a trade-off between library effectiveness and library complexity. We developed a multiplexed, high-throughput screening strategy based on an optimized AsCpf1 nuclease that minimizes library size without sacrificing gene targeting efficiency. Our AsCpf1-based multiplexed library performed similarly well compared to currently available CRISPR/Cas9 libraries, but with a single polycistronic crRNA clone targeting each gene. With this strategy, we constructed the smallest whole-genome knock-out library available, \"Mini-human\" for the human genome, which is one-fourth the size of the smallest CRISPR library currently available.
- organism: human
- assay: CRISPR screen
- research focus: perturbation response prediction
- approach: experimental assay
- Citations
- 44
- FWCI
- 2.32
- Field percentile
- top 10%
Reproducibility ledger
Nobody has reported trying to reproduce this paper yet. Did you replicate an experiment or run the code? Your report saves the next lab a week.
Sign in to file a reproducibility report.
Related papers
- Cas-CLIP: a method for customizing pooled CRISPR libraries
Jiyeon Kweon, Da-eun Kim, … Yongsub Kim·bioRxiv·Feb 25, 2018
- Minimal genome-wide human CRISPR-Cas9 library
Emanuel Gonçalves, Mark Thomas, … Mathew J. Garnett·Genome biologyPeer-reviewed·Jan 21, 2021
- A benchmark comparison of CRISPRn guide-RNA design algorithms and generation of small single and dual-targeting libraries to boost screening efficiency
Sebastian Łukasiak, Alex T. Kalinka, … David Walter·BMC GenomicsPeer-reviewed·Feb 26, 2025
- Improved vectors and genome-wide libraries for CRISPR screening
Neville E Sanjana, Ophir Shalem, Feng Zhang·bioRxiv·Jun 28, 2014
- Minimized double guide RNA libraries enable scale-limited CRISPR/Cas9 screens
Elin Madli Peets, Luca Crepaldi, … Leopold Parts·bioRxiv·Nov 29, 2019
- Optimization of genomewide CRISPR screens using AsCas12a and multi-guide arrays
Sakina M. Petiwala, Apexa Modi, … Daniel Verduzco·The CRISPR JournalPeer-reviewed·Feb 1, 2023