Custom Pooled sgRNA Libraries

The Custom CRISPR Library Service of our partner Cellecta ensures production of a quality library that meets your experimental needs

There are many technical challenges that need to be overcome to produce sgRNA libraries with representative, evenly distributed elements that are amenable to Next-Generation Sequencing (NGS) read-outs.
Our well experienced partner Cellecta offers both, ready-to-use, off-the-shelf Genome-Wide CRISPR libraries, as well as custom-designed CRISPR sgRNA Pooled libraries targeting virtually any set of genes.
Nucleus Biotech works with you to optimize all aspects of the library for your desired application:There are many technical challenges that need to be overcome to produce sgRNA libraries with representative, evenly distributed elements that are amenable to Next-Generation Sequencing (NGS) read-outs.

Custom pooled sgRNA library service description:

1. Oligo Design and Synthesis
For sgRNAs targeting standard human and mouse protein-coding genes, you need only provide us with your list of targets. We design oligonucleotides encoding sgRNAs to your target genes based on the latest guidelines  (e.g. Doench, et al., Nature 2016) as well as some other features we have optimized based on our screening work, to maximize effectiveness and minimize off-target activity.  In addition, we typically employ our improved sgRNA scaffold structure which incorporates the HEAT modifications unless other designs are preferred.
For more specialized applications, researchers may also provide their own guide sequences, or we can work with you to design specialized sgRNAs to meet your needs

2. Cloning
After the design step, we synthesize and clone the pool of oligos in any of our standard library vectors (See Vector Information) or in a customer-provided vector. We can also include a panel of non-targeting, intron-targeting, non-specific cutting, and lethal sgRNA controls to provide reference standards for use when analyzing screening results.

3. Quality Analysis
Once the library is made, we isolate a few dozen constructs for full-insert Sanger sequencing to confirm the configuration of the sgRNA expression cassette and ensure correct insertion. We also deeply sequence all guide sequences by NGS, to confirm full representation of the oligo pool, and assess distribution.
Libraries that do not meet our standards are remade.

4. Deliverables
We provide 500 µg of the plasmid library and the following information:
– All sequence information on the sgRNA guides and vector
– NGS sgRNA distribution data
– Complete cloning site design
– Primer information for sequencing

Custom sgRNA libraries for CRISPR ko, CRISPRa, and CRISPRi with any guide and tracrRNA sequences desired (standard, HEAT-optimized, 10X capture sequences, etc.) can be ordered.

Please use our Service request form  to contact us for more details or request a quote