ES cell based gene targeting services

 

In the past decade, the application of gene targeting technology in murine embryonic stem (ES) cells has made a profound impact on genetic analysis, allowing researchers to explore gene functions in vivo. Based on traditional ES cell-mediated targeting techniques, TurboKnockout® is a leading technology created by Cyagen which can be used to develop complex gene targeting projects to provide mouse models with accurate genetic modification and no off-target effects in as fast as 6 months.

 

The ES cell-mediated gene targeting method creates genetically engineered mice via homologous recombination in embryonic stem (ES) cells. Our services cover a range of conditional knockout, conventional knockout, and knockin model types.

 

>> ES cell mediated Knockout & Knockin Mice

 

Process

 

The steps in the production of gene-targeted mice as shown below:

 

 

 

 

TurboKnockout® Gene Targeting Service

 

Our proprietary ES cell-mediated TurboKnockout® service can provide you with conditional knockout, reporter knockin, and humanization mouse models in as fast as 6-8 months.

 

This proprietary gene targeting method is highlighted by two innovations:

(1) A super competent ES cell line that generates 100% ESC-derived founder mice rather than chimeras, eliminating the need to screen for germline transmission.

(2) A selection cassette on the targeting construct capable of removing itself after ES cell targeting without the need to breed to Flp deleter mice.

 

These innovations eliminate two generations of breeding, shortening production time by 4-6 months as compared to industry standard. For conditional knockouts, it may be possible to breed founders directly to your tissue-specific Cre mice.

 

>> TurboKnockout® Humanized and Knockin Mice

>> TurboKnockout® Knockout (KO) and Conditional KO (cKO) Mice

 

Comparison of Animal Model Generation Methods

 

 

TurboKnockout®

CRISPR/Cas9 Gene Editing

Traditional Gene Targeting

Turnaround time

6-8 months

5-7 months

10-14 months

Donor species & backgrounds

Mouse: C57BL/6, BALB/c

Mouse: C57BL/6, FVB
Rat: Sprague-Dawley (SD), Long Evans

Mouse: C57BL/6, BALB/c

Approach

Homologous recombination in ESC by Cyagen’s TurboKnockout® technology

CRISPR/Cas9 nuclease mediated gene targeting by pronuclear injection

Homologous recombination in ESC by traditional technology

Applications

Knockout (KO)
Conditional knockout (cKO)
Point mutation
Reporter knockin (KI)
ROSA26 knockin (KI)
Humanization

Knockout (KO)
Conditional knockout (cKO)
Point Mutation
Limited knockin (KI)
fragment size

Knockout (KO)
Conditional knockout (cKO)
Point mutation
Reporter knockin (KI)
ROSA26 knockin (KI)
Humanization

Knock-in fragment size limits

 ~20 kb

 

*RMCE Humanization: ~ 300 kb

Up to 15 kb (endogenous)

 ~20 kb

 

*RMCE Humanization: ~ 300 kb

 

Key Features

100% ESC-derived F0

Rapid turnaround

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Potential off-target effects

No

Yes

No

Self-deleting selection cassette

Yes
No need to breed to Flp deleter mice

---

No
Requires breeding to Flp deleter mice

Screening method

PCR + Southern blot

PCR + Sequencing

PCR + Southern blot

Patent Risks

N/A

High-risk

N/A

*Per round of gene targeting

 

Why Choose Cyagen?

  • 15+ years' experience in custom animal model generation
  • 78,000+ animal models generated and delivered worldwide
  • 6,200+ citations in SCI journals
  • 3,000+ university and company partners
  • 100% money-back guarantee option available for most services
  • AAALAC-Accredited and OLAW Assured