High-throughput screening of a CRISPR/Cas9 library for functional genomics in human cells

Nature ◽  
2014 ◽  
Vol 509 (7501) ◽  
pp. 487-491 ◽  
Author(s):  
Yuexin Zhou ◽  
Shiyou Zhu ◽  
Changzu Cai ◽  
Pengfei Yuan ◽  
Chunmei Li ◽  
...  
Keyword(s):  
Functional Genomics ◽  
High Throughput ◽  
High Throughput Screening ◽  
Human Cells

scholarly journals A Perspective on the Future of High-Throughput RNAi Screening

2015 ◽  
Vol 20 (8) ◽  
pp. 1040-1051 ◽  
Author(s):  
Jessica Taylor ◽  
Simon Woodcock
Keyword(s):  
Functional Genomics ◽  
High Throughput ◽  
High Throughput Screening ◽  
Drug Targets ◽  
Gene Knockout ◽  
Strand Break ◽  
Loss Of Function ◽  
Rnai Screening ◽  
Guide Rna ◽  
Cas A

For more than a decade, RNA interference (RNAi) has brought about an entirely new approach to functional genomics screening. Enabling high-throughput loss-of-function (LOF) screens against the human genome, identifying new drug targets, and significantly advancing experimental biology, RNAi is a fast, flexible technology that is compatible with existing high-throughput systems and processes; however, the recent advent of clustered regularly interspaced palindromic repeats (CRISPR)-Cas, a powerful new precise genome-editing (PGE) technology, has opened up vast possibilities for functional genomics. CRISPR-Cas is novel in its simplicity: one piece of easily engineered guide RNA (gRNA) is used to target a gene sequence, and Cas9 expression is required in the cells. The targeted double-strand break introduced by the gRNA–Cas9 complex is highly effective at removing gene expression compared to RNAi. Together with the reduced cost and complexity of CRISPR-Cas, there is the realistic opportunity to use PGE to screen for phenotypic effects in a total gene knockout background. This review summarizes the exciting development of CRISPR-Cas as a high-throughput screening tool, comparing its future potential to that of well-established RNAi screening techniques, and highlighting future challenges and opportunities within these disciplines. We conclude that the two technologies actually complement rather than compete with each other, enabling greater understanding of the genome in relation to drug discovery.

scholarly journals High-Throughput Silencing Using the CRISPR-Cas9 System

2015 ◽  
Vol 20 (8) ◽  
pp. 1027-1039 ◽  
Author(s):  
Mark Wade
Keyword(s):  
Functional Genomics ◽  
High Throughput ◽  
High Throughput Screening ◽  
Signaling Cascades ◽  
Gene Correction ◽  
Pooling Strategies ◽  
Rapid Generation ◽  
Interfering Rna ◽  
Multiple Signaling ◽  
Target Effects

The clustered regularly interspaced short palindromic repeats (CRISPR)/Cas system has been seized upon with a fervor enjoyed previously by small interfering RNA (siRNA) and short hairpin RNA (shRNA) technologies and has enormous potential for high-throughput functional genomics studies. The decision to use this approach must be balanced with respect to adoption of existing platforms versus awaiting the development of more “mature” next-generation systems. Here, experience from siRNA and shRNA screening plays an important role, as issues such as targeting efficiency, pooling strategies, and off-target effects with those technologies are already framing debates in the CRISPR field. CRISPR/Cas can be exploited not only to knockout genes but also to up- or down-regulate gene transcription—in some cases in a multiplex fashion. This provides a powerful tool for studying the interaction among multiple signaling cascades in the same genetic background. Furthermore, the documented success of CRISPR/Cas-mediated gene correction (or the corollary, introduction of disease-specific mutations) provides proof of concept for the rapid generation of isogenic cell lines for high-throughput screening. In this review, the advantages and limitations of CRISPR/Cas are discussed and current and future applications are highlighted. It is envisaged that complementarities between CRISPR, siRNA, and shRNA will ensure that all three technologies remain critical to the success of future functional genomics projects.

A high-throughput screening method based on stably transformed human cells was used to determine the immunotoxic effects of fluoranthene and other PAHs

2008 ◽  
Vol 22 (5) ◽  
pp. 1301-1310 ◽  
Author(s):  
Gertie Janneke Oostingh ◽  
Maria Schmittner ◽  
Angela Karoline Ehart ◽  
Ulrike Tischler ◽  
Albert Duschl
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Screening Method ◽  
Human Cells

Combined High Throughput Screening with QSAR Analysis Unravel Potential Glyoxalase-I inhibitors

2020 ◽  
Vol 16 ◽  
Author(s):  
Mahmoud A. Al-Sha’er ◽  
Qosay A. Al-Balas ◽  
Mohammad A. Hassan
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
3D Qsar ◽  
Human Cells ◽  
Glyoxalase I ◽  
Glyoxalase System ◽  
Qsar Modeling ◽  
Qsar Analysis ◽  
Ubiquitous System ◽  
Further Development

Introduction: Glyoxalase system is ubiquitous system in human cells which has been examined thoroughly for its role in cancerous diseases. It detoxifies endogenous harmful metabolites, mainly methylglyoxal (MG) into non-toxic bystanders. In previous work, our group has explored a series of compounds against glyoxalase I protein. Method: In this research, highthroughput screening approach was used to investigate the activity of in-house database composed of 205 compounds. Results: 15 compounds were found active as glyoxalase I inhibitors. Structure based model Hypo(2ZA0_2_02) combined with 3D-QSAR modeling were applied to predict glyoxalase I inhibition and to explain their activity. The 15 candidates showed more than 50% inhibition with low micromolar IC50 ranges between 5.0 to 42.0 µM. Conclusion: The compounds have been successfully mapped and fitted the Hypo(2ZA0_2_02) model which explain the presence of anti-glyoxalase I activity. This model could be used in future for further development of new and novel glyoxylase I inhibitors.

scholarly journals High-throughput screening for genes that prevent excess DNA replication in human cells and for molecules that inhibit them

Methods ◽  
2012 ◽  
Vol 57 (2) ◽  
pp. 234-248 ◽  
Author(s):  
Chrissie Y. Lee ◽  
Ronald L. Johnson ◽  
Jennifer Wichterman-Kouznetsova ◽  
Rajarshi Guha ◽  
Marc Ferrer ◽  
...  
Keyword(s):  
Dna Replication ◽  
High Throughput ◽  
High Throughput Screening ◽  
Human Cells

In vivo functional genomics of Pseudomonas aeruginosa for high-throughput screening of new virulence factors and antibacterial targets

2003 ◽  
Vol 5 (12) ◽  
pp. 1294-1308 ◽  
Author(s):  
Eric Potvin ◽  
Dario E. Lehoux ◽  
Irena Kukavica-Ibrulj ◽  
Karine L. Richard ◽  
Fran��ois Sanschagrin ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Functional Genomics ◽  
Virulence Factors ◽  
High Throughput ◽  
High Throughput Screening
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