![]() If you use this method in your research, please cite: ![]() Reference genome spiked with gRNA sequences Links to raw data archives and related resources. While the Addgene team is still expanding our plasmid feel free to contact us directly)Please visit our lab's Addgene page for future updates, such as CROP-seq plasmids with fluorescent markers for in vivo and ex vivo use. Derived from one of the most popular constructs for pooled CRISPR screening (LentiGuide-Puro), it retains compatibilty with established, one-step library cloning methods.ĬROPseq-Guide-Puro is now available from Addgene ( 86708. We placed the gRNA expression cassette directly into the 3'LTR, so that the antibiotic resistance mRNA acts as a vehicle to detect gRNAs in single-cell transcriptome data. Our CROPseq-Guide-Puro vector has been designed to make CRISPR gRNAs detectable by single-cell RNA-seq. The protocol contains a detailed listing of all required reagents and oligonucleotide sequences.įor each of the major steps, we present and discuss the expected results. Lentivirus production and cell culture for CROP-seq screens.Pooled, amplification-free cloning of gRNA libraries.DateĬROP-seq step-by-step protocol (Version: 8)ĭetailed description of the CROP-seq method, separated into three chapters: Updated protocols for CROP-seq and links to wet lab reagents. If you are interested in our technology, we would like to hear from you: journal subscription? Read it online for free or ask us to send you the PDF. It is accompanied by a complete step-by-step protocol and software to help interested laboratories get started using CROP-seq. We are excited to share our full article on CROP-seq, which was published on the Nature Methods website today ( DOI: 10.1038/nmeth.4177) and will be part of the March print issue. News and updates DateĬROP-seq paper published in Nature Methods CROP-seq allows pooled CRISPR screens with single-cell resolution, and establishes a paradigm for pooled screening in complex cell populations and differentiation hierarchies. Single-cell transcriptomes can be grouped by the targeted gene or detected gRNA to obtain high-quality gene expression profiles for each perturbation.
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