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The General Introduction of CRISPR Libraries Construction

CRISPR technology has become one of the most effective methods for gene/genome editing. CRISPR libraries/sgRNA libraries can simultaneously edit multiple genes in a signal pathway or an entire genome. This makes it a powerful research tool in CRISPR screening. Through functional screening and enrichment, PCR amplification, and deep sequencing analysis, the functional gene can be identified. With the same techniques, the related signal pathway and molecular mechanism can also be recognized. This technology has been widely used in physiology mechanism, cancer diagnosis, drug discovery, etc.

Synbio Technologies has more than ten years of high standard CRISPR gRNA library experience. We provide customers around the world with precise, efficient, and customized solutions. Some of these include sgRNA design, oligo pool synthesis, CRISPR library/sgRNA library construction, NGS validation, and high-content screening.

Highlights

Precision Editing with Minimal Off-target Effect
One Stop Solution, Starting from Genome Wide sgRNA Design to Library Construction
Construction of Any Type of Library for Any Species
Fast Delivery of Transfection Grade Plasmids

Service Details

Service Specifications	Service Steps	Turnaround Time
sgRNA Design	sgRNA sequences are designed for specific genes 3-6 sgRNAs for each target gene	1-2 weeks
Oligo Pool Synthesis	All designed sgRNAs and negative controls will be synthesized simultaneously	3-4 weeks
sgRNA Library Construction	Cloning sgRNAs pool onto the target vector	2-3 weeks
Plasmid Purification	Transfection-grade plasmid preparation to meet the shipping need	1 week
Library Quality NGS Verification(Optional)	1.100x greater library coverage 2.>70% cloning accuracy rate 3.>99% library accuracy rate by NGS verification 4.<10 library uniformity index	2-3 weeks
Total		9-13 weeks

QUOTE

Workflow

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Case Study

Screening resistance target genes for a drug with identified function

Methods

1.The function and lethal dose of the drug have already been identified.

2.The CRISPR library was synthesized and transfected into cells using lentivirus vector.

3.Screen the drug of high concentration acting on the mixed cell clone continuously over a short period. Then rescreen the surviving cells. These cells are considered to be resistant to this drug. The reason why these cells gain a resistance to this drug is due to mutagenesis.

4.Perform NGS and bioinformatics analysis on the surviving cells, analyze information about the gRNA, and then identify the genes involved in conferring drug resistance.

Results

1.Identification of 6 genes involved in PLX resistant when mutant

2.Identification of NF2 as the resistant target of PLX in melanoma

FAQs

Can any species be designed?

Most common species used with CRISPR technology can be designed. If a desired species is not listed, please reach out!

What is delivered after the library is constructed?

The delivery contains about 200 ug plasmids.

What experiments can synthetic sgRNA libraries be used for?

Some experiments that use synthetic sgRNA libraries include functional gene screening, Tumor Research, Cancer diagnosis, Drug discovery, and many more.

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