CRISPR-sgRNA-library

Pooled CRISPR single guide RNA libraries (sgRNA libraries), are ideal for high-throughput screening of important molecular targets. These libraries leverage the efficiency and specificity of the CRISPR gene editing technology to either knock-out gene expression or transcriptionally activate genes in the genome.

EdiGene offers CRISPR sgRNA libraries for high-throughput knockout of human genes in specific or custom gene groups or pathways. Loss-of-function screening by gene knockout is a powerful tool for systematic genetic analysis in mammalian cells, facilitating gene discovery, genome-scale functional interrogation (e.g. signal transduction pathways) and drug discovery (e.g. target identification and drug mechanism studies).

· sgRNA library

EdiGene’s whole genome sgRNA library targets about 20,000 genes, each gene has 8-10 sgRNAs. Based on annotated function, localization, etc.

Design description:

We designed the human genes knock-out sgRNAs library targeting 18,513 genes annotated in the USCS hg38 database. All potential sgRNAs targeting the CDS region were identified for all the protein coding genes. Then, sgRNAs with a GC content higher than 80% or lower than 20% were removed. Only sgRNAs targeting the first 50% part of the CDS were kept. Those remaining sgRNAs were mapped to the human genome and those had no off-target site were kept as the candidates. We ranked the candidate sgRNAs by the algorithm published by John Doench et.al. in Nature Biotechnology. And the top 10 sgRNAs were selected as the final library. The final library contained 170,815 sgRNAs, and was divided into 10 sub-libraries based on the gene location.

Benefit:

· We optimized the library according to the published data to insure that the sgRNAs had higher efficiency in genome editing.

· 98.7% of the genes in our library have 6 or more sgRNAs, and will give a more robust result for CRISPR screen.

· We supply different sub-libraries, which is smaller and less time consuming for specific usage.

Catalog

Products Name

Target Genes

Price

LA001

EdiGene Human Whole Genome sgRNA Library

19277

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LA002

EdiGene Human Cytoplasm sgRNA Library

6011

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LA003

EdiGene Human Extracellular Space sgRNA Library

1699

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LA004

EdiGene Human Nucleus sgRNA Library

4130

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LA005

EdiGene Human Plasma Membrane sgRNA Library

3279

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LA006

EdiGene Human Other sgRNA Library

4072

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· LncRNA library

Eukaryotic genomes contain both protein-coding and non-coding regions. Transcriptome analyses reveal that up to 90% of the human genome is transcribed into non-coding RNA. Non-coding RNA is further subdivided according to its size into small non-coding RNAs (<200nt) and long non-coding RNAs. The lncRNAs are associated with different molecular and cellular functions and have been associated with cancer, development and diseases. However, the cellular function for most of these newly identified lncRNAs have yet to be elucidated. Also, lncRNAs may play an important part in gene regulation through different mechanisms, including chromatin modification, transcription, post-transcription, interaction with RNA-binding proteins, co-activation of transcription factors and repression of promoters. Based on its proprietary paired gRNA (pgRNA) library design strategy, EdiGene has constructed pgRNA library targeting about 700 lncRNAs3, which will enable researchers to identify molecular functions of IncRNAs in cancer.

 

  1. Zhou Y, et al. (2014) High-throughput screening of a CRISPR/Cas9 library for functional genomics in human cells. Nature 509 : 487.

  2. Peng J, et al. (2015) High-throughput screens in mammalian cells using the CRISPR-Cas9 system. FEBS J. 282 : 2089.

  3. Zhu S, et al. (2016) Genome-scale deletion screening of human long non-coding RNAs using a paired-guide RNA CRISPR–Cas9 library. Nat. Biotechnol. 34 : 1279.