Application of CRISPR-Cas9 technology in the screening of tumor resistance genes:A review

Drug resistance is a major problem faced by tumor cell-targeted drugs.Currently,functional gene screening is the most common strategy for screening drug resistance genes.In recent years,Crispr-cas9 gene editing technology has been widely used in the functional studies of tumor-related genes due to their characteristics of accuracy,simplicity and efficiency.The principle of CRISPR-Cas9 Library Screening Technology and its application in functional Gene Screening are reviewed.At the same time,the application prospect of the Crispr-Cas9 technology is forecasted.

Applications of CRISPR-Cas9 mediated genome engineering

Targeted mutagenesis based on homologous recombination has been a powerful tool for understanding the mechanisms underlying development, normal physiology, and disease. A recent breakthrough in genome engineering technology based on the class of RNA-guided endonucleases, such as clustered regularly interspaced short palindromic repeats(CRISPR)-associated Cas9, is further revolutionizing biology and medical studies. The simplicity of the CRISPR-Cas9 system has enabled its widespread applications in generating germline animal models, somatic genome engineering, and functional genomic screening and in treating genetic and infectious diseases. This technology will likely be used in all fields of biomedicine, ranging from basic research to human gene therapy.

A Comparative Analysis of CRISPR Screening Technologies

This paper offers a general review and comparative analysis of various types of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) technologies. It evaluates the strengths and weaknesses of these technologies to identify the optimal approach for conducting genetic screens. Through an extensive literature review, this paper examines CRISPR nuclease, CRISPR activation (CRISPRa), and CRISPR interference (CRISPRi) screens. This study concludes that CRISPRa and CRISPRi are more advantageous due to their use of deactivated Cas9 proteins that only over-express or deactivate genes rather than irreversibly breaking genes like CRISPRn. Notably, CRISPRa is unique in its ability to over-express genes, while the other two technologies deactivate genes. Future studies may focus on inducing multiple mutations simultaneously—both gain-of-function and gene knockout—to carry out a more complete screen that can test the combinatorial effect of genes. Likewise, targeting both exons and introns can offer a more thorough understanding of a specific phenotype.

Interrogating genome function using CRISPR tools: a narrative review

Genome editing serves as a powerful approach to interrogate the functions of both coding and noncoding sequences.In particular,clustered regularly interspaced short palindromic repeat(CRISPR)/CRISPR associated protein 9(Cas9)system-based editing tools have revolutionized the way we study genome function in mammalian cells,and are being widely used for interrogating critical genes and DNA elements essential for many biological processes.Here,we review CRISPR/Cas9-based genetic tools with an emphasis on CRISPR-mediated high throughput genetic screens in the mammalian genome.