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New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

Posted by Alyssa Cecchetelli on Oct 4, 2018 8:44:18 AM

CRISPR pooled libraries have allowed scientists to easily perform genome-wide screens to effectively and efficiently investigate gene function. CRISPR libraries can be used to knock out, inhibit or activate target genes by combining specific sgRNAs with Cas9 or Cas9 derivatives.

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Topics: CRISPR, CRISPR Pooled Libraries

Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design

Posted by Beth Kenkel on Sep 13, 2018 9:55:58 AM

Genome-wide CRISPR/Cas9 screens are a high-throughput systematic approach for identifying genes involved in a biological process. These screens provide an alternative to genome-wide RNAi screens, which although highly effective, are affected by low on-target efficacy, non-specific toxicity, and off-target effects. The flaws of RNAi screens are well characterized and strategies exist to control for these faults. However, it’s still unclear if similar pitfalls exist for CRISPR screens and how best to design these screens to controls for flaws. Recently the Bassik Lab at Stanford developed a new genome-wide CRISPR knockout screen to analyze the following unanswered questions about CRISPR screen design.

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Topics: CRISPR, CRISPR Pooled Libraries

New Tool for Lineage Tracing: The ClonTracer Library

Posted by Tyler Ford on Sep 22, 2015 10:30:00 AM

This article is based on an interview with Novartis researcher, Carrie Bhang.

The ClonTracer Library, deposited by Carrie Bhang, a research investigator in the In Vivo Pharmacology group at Novartis Oncology, is an exciting new tool that allows researchers to individually label millions of mammalian cells through lentiviral infection and to monitor their abundance and clonal dynamics over time using next generation sequencing (NGS). The library was developed when Carrie was a post-doc in Frank Stegmeier’s lab in Novartis Oncology. 

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Topics: Interview, Viral Vectors, CRISPR Pooled Libraries, Cancer

Genome-wide Screening Using CRISPR

Posted by Joel McDade on Aug 18, 2015 10:30:00 AM

This post was updated on Dec 8, 2017.

What genes are important in your phenotype of interest? Many scientists study diseases for which the underlying genetic cause is not entirely known. Identifying which genes are important for a phenotype can lead to a wealth of additional experiments investigating the role of individual genes or entire pathways in a particular disease process. While CRISPR is certainly not the first means to carry out so-called “forward genetic screening experiments”, it is certainly the most robust. In this blog post, we will discuss how CRISPR libraries are being used to perform genome-wide screens and highlight some of the reagents that have been made publicly available through Addgene. 

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Topics: CRISPR, CRISPR Pooled Libraries

Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

Posted by Kendall Morgan on Feb 24, 2015 2:50:00 PM

CRISPR technology has changed how scientists edit and control genes, but according to the Broad Institute's Silvana Konermann, the first generation of CRISPR-Cas9 plasmids were not designed with gene activation in mind. “We had not managed to create a system to allow us to reliably activate essentially any gene,” she says. The technical leap from mutating and deactivating a gene or genes to selectively activating them with the CRISPR system was a large one.  The question for her then was this: Can you engineer CRISPR-Cas9 activators that work well enough on any gene that they could be used by people with little bioengineering expertise?

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Topics: Plasmid Technology, CRISPR, CRISPR Pooled Libraries

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