CRISPR library complete solution

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CRISPR library is a high-throughput gene screening method established based on CRISPR/Cas9 technology. It identifies phenotype-related genes or screens new drug targets through functional screening, enrichment and deep sequencing analysis. The scope of screening can be the whole genome, a certain gene family, or a certain signal pathway gene. CRISPR library screening has become the preferred platform for large-scale gene function screening benefited from the characteristics of CRISPR/Cas9 such as versatility, low noise, high knockout efficiency, and less off-target effect.

Advantages of Ubigene

Red Cotton CRISPR gene editing system The exclusive data processing system can automatically design 3-6 gRNAs for each gene.
Exclusive CRISPR-U™ technology Gene-editing efficiency elevate 10-20 times due to higher cleavage efficiency and homologous-recombination efficiency.
More than 5000 successful gene-editing cases gene editing has been successfully implemented on more than 200 kinds of cell line. And more than 3000 KO cell lines and 10000 gRNA plasmids have been launched.
Nearly a hundred Cas9 stable cell lines Low cell passage, high cell viability and good cell condition of these cell lines with STR authentication and Cas9 expression detection reports easily accelerate the construction of CRISPR library.

Screening Library Customization

Focusing on gene editing and having rich experience in cell line gene editing, Ubigene can provide 8 major libraries and one-stop services including sgRNA library construction, virus packaging, cell transfection, antibiotic screening, NGS sequencing, and data analysis.

Service Types

Ubigene provides one-stop services for CRISPR-KO, CRISPRa and CRISPRi customized libraries, including sgRNA library construction, virus packaging, cell transfection, antibiotic screening, high-throughput sequencing and data analysis. We offer multiple deliverables meet different research needs.

CRISPR-KO library
Cas9:gRNA complex
The gRNAs of this kind of library mainly target the 5' end exons of coding genes. DNA double strand breaks are caused by Cas9 protein cleavage, and knockout is achieved by introducing frameshift mutations through the non homologous end joining (NHEJ) DNA repair mechanism.
CRISPRa library
dCas9-SAM system
This kind of library can target the transcription-regulatory regions of coding or non-coding genes, achieve the co-expression of Cas9 protein and MS2-P65-HSF1 activation auxiliary protein by connecting dCas9 protein and transcription activator VP64, effectively upregulate the expression level of the genes, and complete high-throughput gain-of-function screening.
CRISPRi library
dCas9-KRAB
The gRNAs of this kind of library mainly target non-coding genes and essential genes (knockout lethality). Gene knockdown was achieved by co-transfecting the gRNA targeting the upstream regulatory region of the gene and a protein of the catalytically inactive dCas9 fused with the transcription repressor KRAB (Kruppel-related frame domain).

Services Details

Service
CRISPR Library Construction
Detail

Including gRNA design, chip synthesis, plasmid construction, plasmid extraction and NGS validation (coverage>99%, uniformity < 10)

Deliverable

Plasmid (100μg, transfection ready) NGS validation report

Price & turnaround
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Amplification of Pre-made Library
Detail

If a pre-made CRISPR plasmid library is provided, we can amplify the library, transform the plasmid to E. coli (coverage>100X), provide NGS validation (coverage 100X-500X), and then deliver the amplified plasmid library for downstream screening.

Deliverable

Small pack (100μg, transfection ready) Large pack (100μg, transfection ready) NGS validation report

Price & turnaround
Inquiry >
Lentivirus Packaging
Detail

3rd generation lentiviral packaging system, highly secure. Lentivirus titer ≥1x10^8 TU/ml guaranteed.

Deliverable

Small pack (1x10^8 TU lentiviral particles) Large pack (5x10^8 TU lentiviral particles) Lentivirus titer test report.

Price & turnaround
Inquiry >
Cell Pool Generation
200+ types of cell lines
Detail

Infect cells with lentivirus library (MOI<0.3, only one virus transduce into one cell), and then conduct antibiotic screening according to the resistance gene carried on the plasmid to generate a cell pool.

Deliverable

Cell pool & NGS analysis report

Price & turnaround
Inquiry >
Functional Screening & NGS Analysis
Detail

Perform cell screening with the compound provided by customers, collect baseline/NC/sample, and extract DNA for NGS and gRNA difference analysis.

Deliverable

Cell pool & NGS analysis report

Price & turnaround
Inquiry >

Technical Process

CRISPR-KO technical process
Library design High throughput automated design of 3-6 gRNAs/gene by the Red Cotton CRISPR gene editing designer.
Chip synthesis sgRNA pool was synthesized by chip synthesis method, and then amplified by PCR.
Plasmid construction 3.To construct a gRNA plasmid library, the sgRNA pool is assembled into the CRISPR-U™ knockout plasmid backbone by Gibson method, followed with transformation, plasmid extraction, and NGS testing.
Lentivirus packaging 4.The gRNA plasmid library is packed with lentivirus (Ubigene used the third generation packaging system, refined the best plasmids ratio, high safety and high titer), and followed with the virus titer testing.
Cell transfection 5.Single-plasmid system can be directly transfected into wild-type cells. But if you use a dual-plasmid system, you need to transfect the gRNA lentivirus library on a Cas9 stable cell line (Ubigene has more than a hundred Cas9 stable cell lines in stock). After cell transfection, antibiotics screening is carried out according to the resistance genes carried on the plasmid backbone, and the successfully transfected cell pool is obtained.
High throughput screening 6.according to different research purposes, select appropriate screening strategies, such as dosing treatment, then conduct positive screening or negative screening to discover candidate genes.
Gene function verification 7.knockout a candidate gene by CRISPR/Cas9 to verify gene function.

Libraries in Stock

Ubigene's off-shelf whole genome CRISPR library of human and mouse can achieve high throughput and rapid screening of functional genes of the entire genome. The human whole genome CRISPR libraries include the single plasmid system (gRNA and Cas9 are in the same plasmid) and the dual plasmid system (gRNA and Cas9 are in different plasmids). At present, only single plasmid system of the mouse whole genome CRISPR library is provided. These three libraries are divided into two sub-libraries, A and B. Each library has designed three gRNAs for each gene, and also contains 1000 control gRNAs for non targeted genomes. Among them, library A has designed four gRNAs for each miRNA. The two sub-libraries can be used separately or together.

EZ-editor™ Human GeCKO v2 genome-wide library Single-plasmid system A/B
Target genes 19,050
gRNA knockout plasmid 122,417
Non-target control sgRNA 1000
EZ-editor™ Human GeCKO v2 genome-wide library Dual-plasmid system A/B
Target genes 19,050
gRNA knockout plasmid 122,417
Non-target control sgRNA 1000
EZ-editor™ Mouse GeCKO v2 genome-wide library Single-plasmid system A/B
Target genes 20,611
gRNA knockout plasmid 130,209
Non-target control sgRNA 1000
Deliverable: 100ug plasmid (virus packaging-ready) QC: Coverage rate > 99%, uniformity < 10 Product details and prices: Request a quote>>

Applications of CRISPR Library

Screening functional genes of tumor

To screen the genes necessary for tumor growth and find new targets for tumor therapy.

To screen gene pairs with synthetic lethality for tumor therapy

To screen the genes causing drug resistance of tumors and study the mechanism of drug resistance

Viral infection related study

To screen host factors related to viral infection, including receptor and endoplasmic reticulum function related genes

Antibody target screening

Use CPRIPR library to validate the specificity of antibody

Signal pathway

Use CPRIPR library to study gene function and its mechanism in signal pathway

Related products

01
Cas9 lentivirus in stock

As low as $900, high titer and viability, efficiently transfect each cell!

02
Cas9 stable cell line in stock

Over a hundred types of cell lines, stably express Cas9, easily achieve gene editing by tranfecting gRNA and donor DNA

03
Lentivirus packaging kit

The 3rd generation packaging system, optimal ratio of different helper plasmids, improve lentivirus yield!


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