# Presentation: 410909

## A mouse for every gene...the Knockout Mouse Program (KOMP) mutagenesis project 

- Kent Lloyd
- University of California, Davis

- NIH/NCRR Council
- January 18, 2007

## The development of murine ES cell technologies in the late 80's marked the emergence of knockout mice as an important animal surrogate for the study of human and animal disease, development, and behavior. 

## Since then, advances in DNA sequencing technologies and throughput have provided complete genome sequences for several commonly used inbred strains of mice: 

- Since then, advances in DNA sequencing technologies and throughput have provided complete genome sequences for several commonly used inbred strains of mice:

- FOR IMMEDIATE RELEASEWednesday, October 25, 2006

**Mouse DNA To Aid Biomedical Research.  **Researchers announced today that they have successfully resequenced the DNA of 15 mouse strains most commonly used in biomedical research. More than 8.3 million single nucleotide polymorphisms (SNPs) were...

## These accomplishments in both knockout technology and genomics have cleared  the way for global efforts to assign function to the 20-25,000 transcribed genes in the mouse genome, by creating “a mouse for every gene”. 

## Over the last 15 years or so, these efforts have taken on a mostly “cottage-industry” approach, which have led to the generation of about 4000 gene-targeted knockouts and 7000 gene traps

## Fall of 2003: NIH convenes a Banbury conference on “Mouse Genome-Wide Targeted Mutagenesis” to discuss completing a knockout library of the mouse genome.

## KOMP

- Nature Genetics 2005;36, 921-924

- 2006-2011

- 2011 and beyond...

## -Supported by 18 Categorical NIH Institutes, 1 Center, 1 Program, and 1 Office

- -Approximately $50million awarded over 5 years
- -4 KOMP RFAs released beween 2005 and 2006

**HG-05-007: The completion of a comprehensive mouse knockout resource**
**DA-06-009: Development and improvement of inbred ES cell lines for use in **
**	****generation of mouse mutants**
**HG-05-008: A data coordination center for the knockout mouse project (KOMP)**
**RR-06-005: Knockout mouse project (KOMP) repository**

## KOMP

**“*****Repatriation*****“...**300 existing mutant lines not publicly available
**“*****Acquisition*****”...**360 selected from Deltagen and Lexicon
** “*****Production*****“...**8000-13,500 DNA constructs, 8000-8500 mutant 	ES cell clones, 500 mutant mice, and cryopreserved embryos

## 2 B6 ES cell grants:  

- -University of Pennsylvania, Philadelphia
- -Samuel Lunenfeld Research Institute of Mount Sinai Hospital in Toronto, Canada

_**1 Informatics grant: **_

- -The Jackson Laboratory, Bar Harbor, Maine

_**1 Distribution repository grant:  **_

- -(pending, Spring 2007)

_**2 Production grants:  **_

- -Children’s Hospital Oakland Research Institute, UC Davis, The Sanger Institute
- -Regeneron, Inc.

## CSD "null first" strategy*

**1**

**2**

**3**

**lacZ-tagged insertion allele**

***FRT***

***FRT***

***loxP***

***loxP***

**lacZ-tagged null allele (********exon)**

**1**

******gal**

**3**

***FRT***

***FRT***

***loxP***

**Cre recombinase**

***based on Testa et al., *****Genesis*****, 2004**

**pre-conditional allele (wild-type)**

**1**

**2**

**3**

**null allele (******** exon****, ****frameshift, NMD)**

**Flp recombinase**

**1**

**3**

_**CSD "null first" strategy***_

**Cre recombinase**

## Regeneron "definitive null" strategy

## it’s a GLOBAL EFFORT

**KOMP**

_**it’s a GLOBAL EFFORT**_

- (“***Nor***th American ***CO***nditional ***M***ouse ***M***utagenesis) Project

- EUCOMM (“***EU***ropean ***CO***nditional ***M***ouse ***M***utagenesis) Programme

## The KOMP gene list: (www.knockoutmouse.org)

*Started with...*
-  **Step 1:** Genes within the NCBI Consensus CDS _[(CCDS) ](http://www.ncbi.nlm.nih.gov/projects/CCDS/)_resource selected:
-  **Step 2:** CCDS genes in MGI with VEGA and Ensembl gene identifiers selected.
-  **Step 3:** Known trapped (_[IGTC](http://www.genetrap.org/)_), targeted, or to be mutated (_[EUCOMM](http://www.eucomm.org/)_) eliminated.
*Continues with...*
**Community invited to nominate their favorite gene** (_[www.knockoutmouse.org](http://www.knockoutmouse.org/)_)...

## after 5 years....

- All mouse genes null
- Many genes conditional-ready
- Collections of both 129Substrain and B6 ES cells
- Germplasm, embryos, ES cells, mice available
- Low cost to obtain
- No reach-through (academic research)
- Less need to use grant money to make mutant mice
- Available to biomedical research community
- Accessible around the world

## KOMP products (mice, ES cells, embryos, data) will enable the study of disease, development, and behavior phenotypes with human homologues, relevant and appropriate to improving and enhancing the translation of basic science research to human (and animal!) patients.... 

*KOMP products (mice, ES cells, embryos, data) will enable the study of disease, development, and behavior phenotypes with human homologues, relevant and appropriate to improving and enhancing the translation of basic science research to human (and animal!) patients.... *