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CRISPR Cas9 sgRNA Library Design and Screening .pdf



Original filename: CRISPR Cas9 sgRNA Library Design and Screening.pdf
Title: CRISPR Cas9 sgRNA Library Design and Screening
Author: http://www.synbio-tech.com/services/crispr-cas9/crispr-cas9-sgrna-library-design/

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CRISPR Cas9 sgRNA Library Design and Screening
CRISPR Cas9 sgRNA Library Design and Screening
Studying gene functions in metabolic pathway or disease development is very
important for disease mechanism research and drug discovery. Leading a new
generation of gene editing technology, the CRISPR-Cas9 system is popular for its
simplicity of operation and its universal applicability. Because of these unique
advantages, CRISPR-Cas9 technology has become a powerful tool for forward
genetic screening. Mutant libraries of various functions can be constructed by
CRISPR, and the genes related to this function then identified through functional
screening and enrichment, PCR amplification, and deep sequencing analysis.
Synbio Technologies provides one-stop gene function screening services based on
the CRISPR-Cas9 sgRNA library system. With our proprietary Syno®3.0 DNA
synthesis platform, we can construct a sgRNA library in a fast and efficient manner.
We can also package the library into lentivirus, transfect cells, perform high
throughput/content screening, and finally identify the target genes by sequencing
and data analysis.

CRISPR Cas9 sgRNA Library Design roadmap

sgRNA library construction and screening workflow
Contact us
Synbio Technologies
1 Deer Park Drive, Suite L-1
Monmouth Junction
NJ, 08852

Tel:+1 732-230-3003
Fax:+1 609 228 5911
Web:www.synbio-tech.com
E-mail:service@synbio-tech.com

Advantages of CRISPR Cas9 sgRNA Library Design
 Proprietary Syno®3.0 DNA synthesis platform: fast and efficient sgRNA
library construction
 Lentivirus transfection system: improve the efficiency of sgRNA transfection
 Multiple screening platform: including cell survival control, immunostaining,
flow cytometry and other screening methods
 One-stop solution: customized services ranging from sgRNA library design,
synthesis, and lentivirus packaging, to high throughput/content screening

CRISPR Cas9 sgRNA Library Design Case Study
Screening resistance targets for a drug with identified function

Methods
1.

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

2. CRISPR-CAS9 gRNA was synthesized and transfected into cells using lentivirus.
3.

Screening: drug of high concentration acting on the mixed cell clone
continuously over a short period, then screening the survived 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 caused by CRISPR-CAS9 gRNA.

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.
Contact us
Synbio Technologies
1 Deer Park Drive, Suite L-1
Monmouth Junction
NJ, 08852

Tel:+1 732-230-3003
Fax:+1 609 228 5911
Web:www.synbio-tech.com
E-mail:service@synbio-tech.com

Result
1.Identification of 6 genes involved in PLX resistant when mutant
2.Identification of NF2 as the resistant target of PLX in melanoma

Service specifications
Synbio Technologies provides one-stop solution including sgRNA library design,
synthesis, lentivirus packaging, and high throughput/content screening
Service Name

Service Specifications

Timeline

Deliverables

CRISPR-Cas9
sgRNA lentivirus
library (Human)

Targeting 21,000 genes;
6 sgRNAs for each gene

Inquiry

Lentivirus,
-80 ℃ preservation

Contact us
Synbio Technologies
1 Deer Park Drive, Suite L-1
Monmouth Junction
NJ, 08852

Tel:+1 732-230-3003
Fax:+1 609 228 5911
Web:www.synbio-tech.com
E-mail:service@synbio-tech.com

customized
CRISPR-Cas9
sgRNA library

Customized gRNA
library, including
genome-scale library,
lncRNA library,
cell apoptosis, cell
proliferation, signal
pathway, ion
channel, nuclear receptor
libraries

Inquiry

Lyophilized
sgRNA plasmid
Sequencing report

lentivirus
packaging of
sgRNA library

Customized sgRNA
design, 6 sgRNA for
each gene
Lentivirus package

Inquiry

Lentivirus

sgRNA library
screening

cell screening using
cell survival
control, immunostaining,
flow cytometry and
other screening methods

Inquiry

Screening report and
sequencing data

Contact us
Synbio Technologies
1 Deer Park Drive, Suite L-1
Monmouth Junction
NJ, 08852

Tel:+1 732-230-3003
Fax:+1 609 228 5911
Web:www.synbio-tech.com
E-mail:service@synbio-tech.com


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