Genome KO Libraries Construction | Synbio Technologies

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General Introduction of Genome KO Libraries Construction

Genome-wide libraries are powerful tools for large-scale gene and genome studies. They enable simultaneous targeting of multiple genes within a pathway or across the entire genome, supporting functional screening, PCR enrichment, and deep sequencing to identify key genes, signaling pathways, and molecular mechanisms. This technology is widely applied in physiology research, cancer studies, and drug discovery.

At Synbio Technologies, we have over ten years of experience in genome-wide library development. Our customized services include library design, oligonucleotide pool synthesis, construction, NGS validation, and high-content functional 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

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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 Genome KO 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 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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