The High Throughout Screening STP

High Throughput Screening : Reagents used to probe gene function

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We have both a human siRNA Smartpool genome library and a corresponding deconvoluted library from Dharmacon, as well as a Mouse Druggable genome collection from Qiagen and a Drosophila genome RNAi library from Ambion. In addition, we have a number of smaller human and mouse siRNA collections themed by gene-function.

Recently the HTS STP has expanded its range of reagents to include CRISPR gene editing technologies. We have developed an arrayed CRISPR screening platform using cell lines expressing inducible Cas9 and transient transfection of synthetic crRNA/tracrRNA. We have a number of arrayed synthetic crRNA collections and are looking to roll out a genome-wide collection soon.

In addition we are continuing to create cell lines ready for CRISPR screening; these lines can obviously also be used for creation of single gene knockouts. We are also continuing to prefect the use of CRISPR related technologies such as CRISPRa and CRISPRi as well as exploring the use of Cas9-variants.

In addition to reagents directly interfering with gene expression, the facility has assembled a collection of 5,500 well-characterised small molecule modulators. These small molecules, many of them drugs currently in clinical use, can be used in assay systems and screens where genetic or RNAi techniques are not possible.

The facility also has additional collections of small molecules that have particular chemical properties and to some extent they have tractable chemistry i.e. they can provide a framework for future elaboration or chemical modification after the primary screen. These include a diversity collection aimed at providing a starting point for proof-of-concept drug development screens and a collection of fragments primarily intended for use in structural studies. They can be used in either cell-based assays or purified proteins.