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CRISPR sgRNA libraries

Introduction

        CRISPR-Cas9 technology is a powerful tool for gene editing that has revolutionized the field of biology. It allows researchers to precisely and efficiently edit DNA at any desired location. CRISPR sgRNA libraries are a type of CRISPR-Cas9 tool that can be used for high-throughput gene screening.

Service process 

Service advantages

        Rich experience in gene synthesis and library construction, ensuring the diversity and uniformity of the library, the library can reach 10^8.

        Select sgRNA expression vectors according to customer needs and provide transfection-grade plasmids.

        Provide a one-stop service from sgRNA design, library construction to lentivirus packaging.

Application cases

        Vemurafenib (PLX), a drug used to treat unresectable or metastatic melanoma with BRAF V600 mutations, can be resistant to the drug due to mutations in cancer cells. Scientists are trying to find the gene targets that lead to resistance to BRAF protein kinase inhibitor vemurafenib (PLX) in melanoma. Using A375 cells as a model, GeCKO forward selection was applied to determine the genes that were knocked out and caused resistance, and to develop drugs against new targets.

A：Timeline of PLX resistance screening for A375 melanoma cells

B：Under the condition of exposure to PLX, the transduced A375 cells stop growing, these cells carry V600E functional gain BRAF mutations

Therefore, a small group of cells that have become resistant through Cas9/sgRNA-mediated editing can be enriched. Vehicle is the control group

After 14 days of PLX treatment, the sgRNA distribution was significantly different from that of the control-treated cells (Wilcoxon rank sum test,

P < 10−10), and it was clustered separately from all other conditions.

The authors found a small set of genes that were enriched with multiple sgRNAs targeting each gene after 14 days of PLX treatment,suggesting that the loss of these specific genes helps A375 cells acquire PLX resistance.

Example figure

Shalem Ophir,Sanjana Neville E,Hartenian Ella et al. Genome-scale CRISPR-Cas9 knockout screening in human cells.[J] .Science, 2014, 343: 84-87.

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