JAX KOMP2 Production Pipeline

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PIs

• Leah Rae Donahue- Director Genetic Resource Science

• Rob Taft- Scientific Director, Reproductive Sciences

• Steve Murray- Research Scientist Genetic Resource Science

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KOMP2 Objectives • Produce mice from 833 IKMC ES cell lines

• Perform quality control (QC)

• Provide mice for phenotyping

• Cryopreserve germplasm

• Develop technology and process improvements that reduce

cost and time of producing mice

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Topics

• Pipeline

• Scale-up

• Innovation

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Pipeline Order ES cells

Import ES cells Quality control

Chimera Production

Germline Testing/Cre Removal

Cryopreservation (conditional ready)

Breeding (het x het)

Viability testing

Fertility testing

Cryopreservation

LacZ staining Distribution for phenotyping

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Staying connected

KOMP Repository JAX – Throughput – Logistics

JAX Production JAX Phenotyping

– Logisitics • Keep phenotyping pipeline full • Minimize breeding costs

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Preparing for expansion

• Space: >30,000 gsf new construction or renovation benefitting KOMP • Systems: New Laboratory Information

Management System

• Staffing: Hiring underway

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Scalability- The JAX repository

0

100

200

300

400

500

600

1 2 3 4 5 6

Num

ber O

f Str

ains

Im

port

ed

Fiscal Year 2010 2005 2009 2008 2006 2007

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Looking ahead

• Phase I – Reduce cost – Improve efficiency

• Phase II

– Many more strains – Limited funding

http://commons.wikimedia.org/wiki/File:NY_429_from_Upper_Mountain_Road_overpass.jpg

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Cost drivers

Acquisition 17%

Chimera production

39%

Breeding 35%

Cryopreservation 9%

Costs

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Use of technology to improve efficiency

0 20 40 60 80

100 120 140 160 180 200

Embryo (flushing)

Embryo (IVF)

Sperm

Day

s

Method

Time

0

20

40

60

80

100

120

Embryo (flushing)

Embryo (IVF)

Sperm Re

lati

ve c

ost

Method

Cost

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Cre driver development

• Deletion activity independent of Cre allele segregation – Being backcrossed to

C57BL/6NJ • Alternative: ROSA26-targeted

CAG-Cre allele – Targeting construct complete – Scheduled to be targeted to

JM8.A3 ES cells late April • Alternative: Import IKMC-

recommended Cre line

X Sox2-Cre♀ Chimera ♂

KO+/-; Cre -

X Cohort breeding

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Improving chimera production

• Inject fewer embryos • Get results faster • Reduce breeding • Chimeras more useful

– Sperm cryopreservation – IVF

• Proof of principle complete • Status: Scaleup

X

+

♂ ♂

XXXX

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Artificial Insemination

• Standard approach in other species

• Historically unreliable • Improved method

– Litter size comparable to natural mating (fresh sperm)

• Value – Reduce breeding time – Reduce breeding cost

Nakagata, N. Exp Anim. 1992;41(4): 519-522.

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Summary

• JAX is prepared to meet the needs of the KOMP2 Reanimation Project – Modular scalable pipeline based on existing

infrastructure – Experienced microinjection group

• JAX is preparing to meet future KOMP2 needs – Infrastructure investment – Technology development

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Questions?