Pooled Screening

In a pooled functional genomic screen, a vast number of perturbations is introduced in a single cell population using a lentiviral library resulting in a mixed pool of cells. Subsequently, this population is challenged with e.g. drug treatments or proliferation and analyzed using Next-Generation Sequencing (NGS) for deconvolution.

This allows to test a large number of gene-edited cell at once in a high-throughput and cost-efficient manner, which is why it is often used for primary whole genome screens.

By identifying common patterns in cellular responses to perturbations, researchers may gain valuable insights into the genetic factors influencing the disease phenotype. Generally pooled screens focus on detecting broad effects rather than elucidating individual gene functions. However recently there are more advanced read-out technologies on the rise like single-cell or in-situ sequencing that allow for more detailed analysis in a pooled screening format.

Advantages:

• Less expensive
• Large number of genes can be integrated at once
• Suitable for longer in vivo studies
• No specialized/automated equipment needed

Read-out methods: 

• NGS At Different Time Points or After Different Selective Pressures
• NGS Combined with Certain Biological Characteristics by FACS Sorting Using Reporter Gene Or Surface Biomarker
• scRNA Sequencing linking gRNA Expression with Transcriptomic Profile
• In-Situ Sequencing linking gRNA Expression with Imaging Phenotype