Target Screening: CRISPR Library vs Traditional Omics

Target Screening: CRISPR Library vs Traditional Omics

Target screening is a crucial task in the fields of life science research and precision medicine. It provides key directions for the diagnosis, treatment, and drug development of diseases. The emergence of CRISPR technology undoubtedly brought revolutionary changes to this field. There are various methods for target screening, including traditional omics methodologies and CRISPR library screening methodologies, but each method has its own advantages and limitations.

Traditional omics methodologies play an important role in drug target screening, mainly including genomics, transcriptomics, proteomics, and metabolomics. These methodologies reveal the changes in biological systems under different physiological or pathological states through large-scale detection and analysis of genes, transcripts, proteins, or metabolites in biological samples, to identify potential disease-related targets.

Figure 1. Traditional omics methodologies for target screening

Genomics:

Focuses on studying the entire genomic information of an organism. For example, genome-wide association studies (GWAS) compares the genome sequences of a large number of cases and control populations to identify single nucleotide polymorphisms (SNP) that are significantly associated with disease traits. These associated sites may reside in coding regions or regulatory regions of genes, thereby affecting the function or expression level of genes and being associated with the occurrence and development of diseases.

Transcriptomics:

Primarily studies the collection of all RNA transcripts transcribed by cells under specific conditions. Through RNA sequencing (RNA-seq) technology, gene expression level information can be obtained to analyze differentially expressed genes between disease tissues and normal tissues.

Proteomics:

The study of protein composition, expression levels, modification states, and interactions between proteins in cells or tissues. Proteomic analysis based on mass spectrometry technology can identify a large number of proteins and quantitatively analyze their differences in different samples.

Metabolomics:

The study of the types, quantities, and variations of metabolites in organisms. By detecting metabolites in samples such as blood, urine, and tissues using techniques such as gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS), it is possible to detect metabolic pathway disorders in disease states.

CRISPR/Cas9 library screening technology is an efficient method for studying gene function. By constructing a large library of guide RNAs (gRNAs) targeting different genes and introducing them into cell populations, the genome is edited using Cas9 protein. Then, through specific screening strategies such as cell survival, proliferation, phenotype changes, etc., genes related to specific biological processes or diseases can be identified.

Figure 2. CRISPR screening workflow

CRISPR Library vs Traditional Omics

Advantages of CRISPR Library Screening

1. In terms of functional verification

The CRISPR library screening technology can directly study the functional loss or acquisition of genes, and clarify the causal relationship of genes in specific biological processes.

2. Comprehensive screening

CRISPR library screening can perform unbiased screening across the entire genome, theoretically identifying any genes related to screening conditions, including some genes that have not been studied or whose functions are unknown.

3. Target operability

Compared to traditional omics methods that screen for targets such as proteins and metabolites, which require complex drug design and development processes (such as developing protein target small molecule inhibitors or antibody drugs), CRISPR library screening identifies targets that are mostly directly editable genes, making it easier for subsequent gene therapy drug development. For example, for single gene inherited disease pathogenic genes, gene editing strategies can be directly developed.

Application: You can click to view our previous posts, CRISPR Library Screening Applications in Research on Acute Leukemia, CRISPR Library | Unlocking New Insights into Breast Cancer Targets, CRISPR Library: Opening a New Perspective for Virus Research - Special Guide and so on. In addition, there are many useful CRISPR Library Application posts available for review. We will also launch more CRISPR library related content in the future. Stay tuned!

Based on the above advantages, more and more researchers are choosing CRISPR libraries to screen for new targets and publish high IF articles! However, CRISPR library screening technology is complex, requiring the construction of high-quality libraries, precise cell transduction, and high-throughput sequencing analysis, relying on professional laboratories and personnel.

Ubigene CRISPR-iScreen™platform can provide one-stop services from high-throughput sgRNA library construction to virus packaging, cell infection, drug screening, high-throughput sequencing, and data analysis. Ubigene CRISPR-iScreen™technology utilizing self-developed high efficiency competent cell and exclusive Cell Pool preparation processes, ensures a Cell Pool library coverage of up to 99%.

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References

[1] Joung JK, et al. Genome - scale CRISPR - Cas9 knockout and transcriptional activation screening. Nat Protoc. 2017;12 (4):828 - 863.

[2] Doench JG, et al. Genome - wide CRISPR - Cas9 knockout screening in human cells. Science. 2016;353 (6304):aaf8729.

[3] Luo B, et al. Genome - wide and pathway - focused CRISPR - Cas9 screens identify mediators of cancer drug resistance. Cell. 2015;161 (6):1247 - 1261.

[4] Reuveni E, et al. Integrating CRISPR screens with multi - omics data to decipher gene regulatory networks. Nat Rev Genet. 2017;18 (7):477 - 490.

[5] Tzelepis K, et al. CRISPR - Cas9 screens in cancer cells to identify novel drug targets and resistance mechanisms. FEBS Lett. 2016;590 (17):2888 - 2901.

[6] Barrera LA, Alvarez-Varela A, Kazane K, et al. CRISPR screens in vivo for tissue-specific cancer drivers. Science. 2016;353 (6307):1160 - 1163.

[7] Tzelepis K, de Stanchina E, D'Arcangelo M, et al. CRISPR-Cas9 screens in cancer cells to identify novel drug targets and resistance mechanisms. FEBS Lett. 2016;590 (17):2888 - 2901.