For researchers looking for better flexibility and higher performance CRISPR screening libraries, IDT’s custom synthetic gRNA libraries offer a highly customizable option, strong CRISPR expertise and support, and an optimized RNA synthesis process.
Custom libraries for better CRISPR screening.
Custom arrayed synthetic guide RNA libraries are a great way to accomplish CRISPR knockout screens. They accelerate your research by avoiding the cloning and sequencing associated with lentiviral screens while also integrating into existing automation workflows. IDT has experience designing and manufacturing arrayed panels of gRNAs in a range of sizes, from a few guides to thousands or more. Our high-capacity manufacturing systems offers delivery of your custom CRISPR panel starting at as fast as five business days.
We know your time is valuable and we’ll prioritize your inquiry. Click on “Request consultation” to provide brief information about your CRISPR library project or interests, and we’ll be in touch to discuss your project ASAP.
Alt-R Custom CRISPR gRNA libraries are available for all CRISPR nucleases, including Cas9, Cas12a, Cas13, prime editing enzymes, and other alternative systems. These libraries were developed to address the need for better CRISPR screening solutions. They are chemically modified guide RNAs synthesized on IDT’s proprietary, high-fidelity RNA manufacturing platform to provide high quality, reliable gRNA libraries with fast delivery.
Features | Options |
---|---|
Design | Predesigned, custom, user-provided |
CRISPR systems | Cas9, Cas12a, Cas13, prime editing, and other alternative systems |
Guaranteed yield | 0.5 nmol, 2 nmol, 5 nmol, and custom normalized deliverables |
Cas9 gRNA formats | 2-part crRNA:tracrRNA complex and sgRNA |
Custom lengths supported | 30–150 nt |
Chemical modifications | 2’-O-methyl RNA, PS linkages, end-blocking Alt-R modifications |
Plate types | 96- & 384-well PCR, Deep-well, V-bottom, Echo, custom options available |
Formulation options | Multi-guide per well; pooled by gene Arrayed (single gRNA/well) Custom formulations upon request |
QC | Individual ESI/MS |
Supporting reagents & functional analysis pipeline (optional) | WT Cas9, HiFi Cas9, Cas12a. and Cas12a Ultra Glycerol-free options available in tubes or plates (ideal for robotics) Electroporation Enhancers rhAmpSeq™ CRISPR Analysis System (NGS-based on-/off-target editing analysis) |
Our Alt-R CRISPR enzymes are available in a variety of formats, with stock sizing available up to 50 mg. Larger formats and lot matching are also available for all products upon request.
Cas protein | Available versions | Key features |
---|---|---|
S.p. Cas9 Nuclease V3 | Wild-type, 50% Glycerol Wild-type, Glycerol-free High Fidelity (HiFi) Wild-type Fluorescent-fusion | Targeting GC-rich regions Low viscosity for high-throughput applications Reduced off-target activity* Fluorescent label for enrichment (GFP or RFP) |
A.s. Cas12a (Cpf1) V3 | Wild-type Ultra | Targeting AT-rich regions Increased on-target activity* |
L.b. Cas12a (Cpf1) | Ultra | Increased on-target activity and low-temperature tolerance* |
* when compared to corresponding Wild-type controls
We investigated the efficiency of inducing non-homologous DNA end joining (NHEJ) by designing 3 sgRNAs per gene (N=95 genes) via an internal pipeline and using IDT’s Alt-R Cas9 nuclease S.p. Cas9 WT V3, electroporation enhancer, and rhAmpSeq™ CRISPR analysis system. The combined use of the Alt-R guides and additional CRISPR products provided an effective, high-throughput editing solution with >90% NHEJ editing in 99% of targeted genes (Figure 1).
Figure 1. A 3-guide-per-gene library design resulted in >90% NHEJ editing in almost all (90/91) targeted amplicons.
95 genes of interest were fed into an internal design pipeline to generate 3 guide RNA designs per gene, all
located within a 500 base span. Once target sites were identified, singleplex genotyping assays for each gene were selected using the rhAmpSeq Design Tool. K562 cells were transfected with 3 sgRNAs complexed to Alt-R S.p. Cas9 WT Nuclease V3 per well
(final concentration = 1 µM per guide). rhAmpSeq CRISPR Library preparation and subsequent sequencing on the Illumina MiSeq platform was then performed. IDT’s rhAmpSeq Analysis Tool revealed a high level of NHEJ-type editing events (dark
blue) in nearly all amplified regions (N = 91 amplicons with sequencing reads >500).
To highlight the editing efficiency of sgRNAs, we designed sgRNAs targeting 255 sites across the human genome and delivered them to Jurkat cells (a human T-lymphocyte-derived cancer cell line) along with Alt-R S.p. WT Cas9 Nuclease V3. The result of this experiment shows that Alt-R CRISPR-Cas9 sgRNAs provide high levels of editing.
Figure 2. High levels of editing with Alt-R CRISPR-Cas9 sgRNAs.
Ribonucleoprotein (RNP) complexes were formed with Alt-R S.p. WT Cas9 Nuclease V3, combined with Alt-R Cas9 sgRNAs synthesized for 255 randomly selected Cas9 guide RNA sites across the human genome. RNP complexes (4 μM) were delivered into Jurkat cells via a Nucleofector™ system (Lonza) in the presence of Alt-R Electroporation Enhancer. Genome editing efficiencies were determined by target amplification followed by next generation sequencing (NGS) on an Illumina™ instrument.
Features | Options |
---|---|
Design | Predesigned, custom, user-provided |
CRISPR systems | Cas9, Cas12a, Cas13, prime editing, and other alternative systems |
Guaranteed Yield | 0.5 nmol, 2 nmol, 5 nmol, and custom normalized deliverables |
Cas9 gRNA formats | 2-part cRNA:tracrRNA complex and sgRNA |
Custom lengths supported | 30-150 nt |
Chemical modifications | 2'-O-methyl RNA, PS linkages, end-blocking Alt-R modifications |
Plate types | 96- & 384-well PCR, Deep-well, V-bottom, ECHO, custom options available |
Formulation options | Multi-guide per well; pooled by gene Arrayed (single gRNA/well) Custom formulations upon request |
QC | Individual ESI/MS |
Supporting reagents & functional analysis pipeline (optional) | WT Cas9, HiFi Cas9, Cas12a, and Cas12a Ultra Glycerol-free options available in tubes or plates (ideal for robotics) Electroporation Enhancers rhAmpSeq™ CRISPR Analysis System (NGS-based on-/off-target editing analysis) |
Yes, we offer predesigned Cas9 CRISPR crRNA and additional options for creating customizable gRNA libraries.
For more information, please visit our Custom guide RNA Libraries page.