Custom Pooled sgRNA Libraries

Cellecta´s Custom CRISPR Lentiviral Pooled sgRNA Library Service: Any target genes, any size and any 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.

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:

Custom pooled lentiviral sgRNA library service description:

1. Oligo Design and Synthesis
Just send us your target gene list, indicate the desired number of sgRNAs per target, and let us know your application. Our partner Cellecta will design oligonucleotides encoding sgRNAs targeting  your genes of interest based on the latest guidelines  (e.g. Doench, et al., Nature 2016) as well as some other features optimized based on Cellecta´s extensive screening work, to maximize effectiveness and minimize off-target activity.  Typically Cellecta´s improved sgRNA scaffold structure which incorporates the HEAT modifications are employed unless other designs are preferred.
For 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, the pool of oligos is evenly synthesized and cloned in any of Cellecta´s high efficiency lentiviral vectors (See Vector Information) or in a customer-provided vector.  Typically a panel of non-targeting, intron-targeting, non-specific cutting, or lethal sgRNA controls is included in the pool to provide reference standards for use when analyzing screening results.

3. Quality Analysis
Once the library is made, a few dozen constructs are analyzed by full-insert Sanger sequencing to confirm the configuration of the sgRNA expression cassette and ensure correct insertion. The entire guide sequences are deeply sequenced by NGS to confirm full sgRNA representation in the oligo pool, and assess distribution of each sgRNA sequence in the library.

4. Deliverables
We provide 500 µg of the plasmid library and the following data:
– Complete sequence information on the sgRNA guides and vector
– NGS QC data files demonstrating the distribution of each sgRNA sequence in the pool
– Cloning site design and sequencing primer information

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