CRISPR sgRNA Control Constructs

Variations of the CRISPR/Cas9 system can be used for cell engineering (e.g., targeted gene knockout, knockdown, or activation), to initiate knock-ins, and for functional genetic screening.

For all of these types of studies, it is often useful to have control constructs that target known genes, essential genes, or are validated not to target any gene (i.e., negative controls). Cellecta offers a range of pre-made sgRNA constructs in various vector backbones to use as controls for CRISPR-related applications.

FAQ's:

1) Do CRISPR libraries include positive and negative controls?

A: Several hundred non-targeting guides are included as negative controls in Cellecta's off-the-shelf, whole-genome libraries. A set of positive control guides is also included for cell viability screens. These controls target essential genes, which makes them toxic to cells. As a result, these guides will be depleted in the next-generation sequencing (NGS) readout from a bulk viability assay.

2) How does the sgPOLR2L control work?

A: sgPOLR2L acts as a positive control for viability assays. Because POLR2L is an essential gene, cells that are successfully targeted or transduced with sgPOLR2L will lose POLR2L function and die.

3) What is the origin of the Cas9 gene?

A: The Cas9 gene Cellecta uses is a synthetically designed, codon-optimized sequence based on S. pyogenes; it is not a natural sequence.