Open Access
High-throughput functional genomics using CRISPR–Cas9
Ophir Shalem,Ophir Shalem,Neville E. Sanjana,Neville E. Sanjana,Feng Zhang,Feng Zhang +5 more
- Vol. 16
TLDR
Recent advances using Cas9 for genome-scale screens are described, including knockout approaches that inactivate genomic loci and strategies that modulate transcriptional activity.Abstract:
Forward genetic screens are powerful tools for the discovery and functional annotation of genetic elements. Recently, the RNA-guided CRISPR (clustered regularly interspaced short palindromic repeat)-associated Cas9 nuclease has been combined with genome-scale guide RNA libraries for unbiased, phenotypic screening. In this Review, we describe recent advances using Cas9 for genome-scale screens, including knockout approaches that inactivate genomic loci and strategies that modulate transcriptional activity. We discuss practical aspects of screen design, provide comparisons with RNA interference (RNAi) screening, and outline future applications and challenges.read more
Citations
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Patent
Novel crispr enzymes and systems
TL;DR: In this article, the authors proposed a method for non-naturally occurring or engineered DNA or RNA-targeting CRISPR effector protein and at least one targeting nucleic acid component like a guide RNA.
Journal ArticleDOI
CRISPR-dCas9 mediated TET1 targeting for selective DNA demethylation at BRCA1 promoter.
TL;DR: A programmable CRISPR-Cas9 based demethylase tool containing the deactivated Cas9 fused to the catalytic domain (CD) of Ten-Eleven Translocation dioxygenase1 (TET1CD) and TET1-dCas9 fusion proteins-mediated demethylation at a target region in BRCA1 gene promoter, a model tumour suppressor gene is examined.
Book ChapterDOI
Next-Generation Sequencing — An Overview of the History, Tools, and “Omic” Applications
TL;DR: The advances, applications, and challenges of NGS are reviewed starting with a history of first-generation sequencing, the bioinformatics issues confronting NGS data stor‐ age and analysis, and the impacts made in the fields of genetics, biology, agriculture, and medicine in the brave, new world of ”omics.
Journal ArticleDOI
Epigenetics in Insects: Genome Regulation and the Generation of Phenotypic Diversity.
TL;DR: This review provides an introduction to the field of molecular epigenetics in insects and provides a brief overview of techniques for profiling and perturbing individual facets of the epigenome.
Journal ArticleDOI
CRISPR library designer (CLD): Software for multispecies design of single guide RNA libraries
Florian Heigwer,Tianzuo Zhan,Tianzuo Zhan,Marco Breinig,Jan Winter,Dirk Brügemann,S Leible,Michael Boutros +7 more
TL;DR: CRISPR library designer (CLD) is described, an integrated bioinformatics application for the design of custom single guide RNA (sgRNA) libraries for all organisms with annotated genomes that predicts a high fraction of functional sgRNAs.
References
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MicroRNAs: Genomics, Biogenesis, Mechanism, and Function
TL;DR: Although they escaped notice until relatively recently, miRNAs comprise one of the more abundant classes of gene regulatory molecules in multicellular organisms and likely influence the output of many protein-coding genes.
Journal ArticleDOI
Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans
Andrew Fire,SiQun Xu,Mary K. Montgomery,Steven A. Kostas,Steven A. Kostas,Samuel E. Driver,Craig C. Mello +6 more
TL;DR: To their surprise, it was found that double-stranded RNA was substantially more effective at producing interference than was either strand individually, arguing against stochiometric interference with endogenous mRNA and suggesting that there could be a catalytic or amplification component in the interference process.
Journal ArticleDOI
A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity.
Martin Jinek,Krzysztof Chylinski,Krzysztof Chylinski,Ines Fonfara,Michael H. Hauer,Jennifer A. Doudna,Emmanuelle Charpentier +6 more
TL;DR: This study reveals a family of endonucleases that use dual-RNAs for site-specific DNA cleavage and highlights the potential to exploit the system for RNA-programmable genome editing.
Journal ArticleDOI
Multiplex Genome Engineering Using CRISPR/Cas Systems
Le Cong,Le Cong,F. Ann Ran,F. Ann Ran,David M. Cox,David M. Cox,Shuailiang Lin,Shuailiang Lin,Robert P. J. Barretto,Naomi Habib,Patrick D. Hsu,Patrick D. Hsu,Xuebing Wu,Wenyan Jiang,Luciano A. Marraffini,Feng Zhang +15 more
TL;DR: The type II prokaryotic CRISPR (clustered regularly interspaced short palindromic repeats)/Cas adaptive immune system has been shown to facilitate RNA-guided site-specific DNA cleavage as discussed by the authors.
Journal ArticleDOI
RNA-Guided Human Genome Engineering via Cas9
Prashant Mali,Luhan Yang,Kevin M. Esvelt,John Aach,Marc Güell,James E. DiCarlo,Julie E. Norville,George M. Church,George M. Church +8 more
TL;DR: The type II bacterial CRISPR system is engineer to function with custom guide RNA (gRNA) in human cells to establish an RNA-guided editing tool for facile, robust, and multiplexable human genome engineering.