Genomic engineering in cell lines is a versatile tool for studying gene function, designing diseases models, biopharmaceutical research, drug discovery and many other applications. CRISPR (Clustered Regulatory Interspaced Short Palindromic Repeats)/Cas9 systems is a newly developed yet the most popular method for genome editing. It has been widely used in current biology, functional genome screening, cell-based human hereditary disease modeling, epigenomic studies and visualization of cellular processes.

1. Stable Knockout Cell Line Generation
info@creative-biogene.com
Website: https://www.creative-biogene.com

2. Brief Introduction
Genomic engineering in cell lines is a versatile tool for studying gene function,
designing diseases models, biopharmaceutical research, drug discovery and
many other applications. CRISPR (Clustered Regulatory Interspaced Short
Palindromic Repeats)/Cas9 systems is a newly developed yet the most popular
method for genome editing. It has been widely used in current biology,
functional genome screening, cell-based human hereditary disease modeling,
epigenomic studies and visualization of cellular processes.

3. Brief Introduction
CRISPR/Cas9 system consists of a “guide” RNA (gRNA)
and a bacterial CRISPR-associated endonuclease (Cas9).
The gRNA is a short synthetic RNA composed of a Cas9-
binding “scaffold” sequence and ∼20 nucleotide
“targeting” sequence that defines the target genomic
site to be modified. Cas9 contains two nuclease domains
to induce site-specific DNA cleavage. It’s a scalable
genome-wide editing technology for its ease of
generating gRNAs. The simplicity and high-efficiency of
CRISPR/Cas9 system make it a preferable genomic
knockout method to the traditional ZFN and TALEN
system. Our scientists are experts at performing gene
knockout with CRISPR/Cas9, from designing gRNA
constructs to transfection and single clone generation of
a wide range of cells, including difficult-to-transfect and
tumor cell lines.

4. 1 Single Gene Knock Out Cell Line
Generation
2 Multiplexed Gene Knock Out Cell
Line Generation
3 Fragment Deleted Stable Cell Line
Generation
CONTENTS

5. LOGO
CRISPR/Cas9 is a revolutionary technology in the field of gene engineering. It has
expedited the course of biopharmaceutical research and drug discovery, by providing
scientists with a versatile tool to knockout any gene, in any cell and without
introducing foreign DNA. The benefits of CRISPR/Cas9 over previous forms of gene
editing, such as TALENs and zinc finger nuclease (ZFN), are that it is much simpler to
implement and has higher efficiency at performing bi-allelic gene modifications.
Creative Biogene is experienced to provide a full CRISPR/Cas9-based single gene
knockout service using any mammalian cell line and targeting any gene. Our
scientists are experts at performing gene knockout with CRISPR/Cas9, from
designing gRNA constructs to transfection and single clone generation of a wide
range of cells, including difficult-to-transfect and tumor cell lines.
Single Gene Knock Out Cell Line Generation

6. LOGO Single Gene Knock Out Cell Line Generation
Our services include:

7. LOGO Multiplexed Gene Knock Out Cell Line Generation
Gene families or biological pathways usually contain several related genes that
collaborate in the same role. Increasing demand in loss-of-function screens has
warranted the need for a multiplexed gene knock out cell line. Multiplexing with
CRISPR/Cas9 can accelerate genome engineering efforts in biopharmaceutical
research and drug discovery.
Creative Biogene provides additional rounds of transformation and selection until
every targeted gene is knocked out. Mammalian cells exposed to two or three
gRNAs in turn resulted in the generation of cell lines harboring double or triple
gene knockouts, that is, the successful disruption of two or three genes. All
multiplexed knockout events were obtained with DNA mutations in targeted genes,
and harbored the expected knockout phenotype(s). Investigation of entire gene
families or biological pathways is enabled by our dual/triple gene knock out cell
line generation service.

8. LOGO
 Biallelic mutation/deletion confirmed by PCR and DNA sequencing.
 KO stability guaranteed up to 20 passages.
 Both products enable unbiased, powerful loss-of-function screens.
 Targeted screening of specific pathways of importance.
 Thorough screening of pathway or disease-focused genes.
 Used as screening tools in the development of drugs.
 To understand the mechanism of action of a drug.
 To target specific biological processes.
Multiplexed Gene Knock Out Cell Line Generation
The KO status of each cell line is thoroughly tested as follows
Applications

9. Multiplexed Gene Knock Out Cell Line Generation Service Case

10. LOGO
CRISPR/Cas9 has been widely utilized to facilitate functional knockout of protein coding
genes. However, a single site of gene editing may not be sufficient to disrupt protein’s
structure (for example, miRNA stem-loop) and function. To overcome this potential
limitation, paired guide RNAs targeting both 5’ and 3’ of the target region have been
shown to result in larger deletions, thus resulting in loss-of-function completely.
Fragment Deleted Stable Cell Line Generation
Dual sgRNA-directed large gene deletion mediated by CRISPR/Cas9 system is a robust
tool for generating deletions of long non-coding RNAs (lncRNAs), MicroRNAs (miRNAs)
and regulatory sequences. Several recent studies have reported the CRISPR/ Cas9
system-induced large genomic deletion, ranged from 23kb to 1Mb, in mouse, C. elegans,
rabbit and human cell lines.

11. LOGO Fragment Deleted Stable Cell Line Generation
Creative Biogene is experienced in producing a fragment deleted cell line using any mammalian cell
line and targeting sequences of your interest. Utilization of two sgRNAs targeting the immediate
sequence upstream and downstream is a viable method for functional knockout of a target protein
completely.

12. LOGO
One-stop service is provided from design of
gRNAs, cell transfection, preparing of single
clones to sequencing analysis, only needing
customers to provide target genes information
and the cell line of interest.
Professional researchers
have rich cell culture experience and
plentiful CRISPR genome editing experience.
Cell lines can be made from nearly
any cell type, including cells that are
difficult to transfect and grow incl. primary cells.
Our CRISPR gene knockout technology
applies to targets of any genes and
mammalian cells.
Advantages

13. THANKS！
info@creative-biogene.com
Website: https://www.creative-biogene.com