Knockout Mouse Production and Cryopreservation (U42) | Apply Online

Frequently Asked Questions (FAQs)

What is the NIH funding opportunity RFA-RM-10-013?

RFA-RM-10-013 is an NIH funding opportunity titled "Knockout Mouse Production and Cryopreservation (U42)." It was designed to expand and operationalize large-scale gene knockout resources being generated by the International Knockout Mouse Consortium by turning existing gene-targeted embryonic stem (ES) cell lines into validated knockout mouse lines, preserving those lines, and supplying mice to NIH-supported phenotyping centers.

What is the main purpose of this program?

The central purpose is to convert gene-targeted ES cell reagents from major international programs into living knockout mouse lines, confirm quality and correct genotypes through quality control (QC), provide those mice to NIH-funded Mouse Phenotyping Centers for standardized phenotyping, and cryopreserve germplasm so the mouse lines can be reliably maintained and distributed long-term.

Where do the starting ES cell lines come from?

The program relies on existing gene-targeted ES cell lines produced by major international resources, notably KOMP and EU-associated resources referenced in the announcement (komp.org and eucomm.org/eummcr.org).

What kinds of activities were applicants expected to propose?

Applications were expected to describe a production-scale, service-oriented operation that: (1) imports knockout ES cell lines from international repositories, (2) performs QC on incoming ES cell materials, (3) generates knockout mice from those ES cells and establishes germline transmission, (4) performs QC on resulting mouse lines and derivatives to confirm the intended genetic alteration, (5) ships knockout mice to NIH-funded phenotyping centers for standardized phenotyping, and (6) cryopreserves germplasm (typically embryos and/or sperm) and sends aliquots to the KOMP Repository for long-term preservation and distribution capability.

Why is quality control (QC) emphasized in this FOA?

QC is emphasized because downstream phenotyping and community use depend on accurate genotypes, reliable gene targeting events, and consistent colony and resource management. The FOA describes QC as critical for ensuring that the produced animals and associated materials meet standards needed for standardized phenotyping and broader biomedical use.

What does "Knockout Mouse Production" mean in this opportunity?

In this FOA, knockout mouse production refers to taking existing gene-targeted ES cell lines and converting them into living knockout mouse lines, for example by generating mice through procedures such as blastocyst injection and breeding to establish germline transmission, followed by validation of the resulting genetic alteration through QC.

What does "Cryopreservation" mean in this opportunity?

Cryopreservation here refers to banking germplasm from produced mouse lines, typically embryos and/or sperm, to ensure long-term preservation, renewable access, and redundancy. The FOA also calls for sending aliquots of cryopreserved materials to the KOMP Repository.

How does this FOA connect to NIH-funded Mouse Phenotyping Centers?

A required operational link in this FOA is coordination with separate NIH-funded Mouse Phenotyping Centers (funded under the companion FOA RFA-RM-10-011). The knockout production sites would ship knockout mice to these centers so standardized phenotyping can be conducted and functional consequences of gene disruption can be measured consistently.

What is the relationship between RFA-RM-10-013 and RFA-RM-10-011?

RFA-RM-10-013 focuses on production, QC, shipping to phenotyping centers, and cryopreservation of knockout mouse lines. RFA-RM-10-011 is described as a companion FOA supporting Mouse Phenotyping Centers that perform standardized phenotyping on the mice generated and delivered through the production pipeline.

What is the relationship between this U42 FOA and the companion U54 announcements?

The FOA notes it operated in parallel with additional companion announcements using the U54 mechanism (RFA-RM-10-012 and RFA-RM-10-011). Together, these announcements reflect a coordinated, multi-site national infrastructure where different awardees handled complementary parts of the pipeline, including production/cryopreservation and phenotyping.

What does it mean that this award uses a U42 cooperative agreement mechanism?

A U42 is a cooperative agreement mechanism, indicating NIH anticipated an active partnership role rather than a fully hands-off grant. Under this structure, the awardee would run day-to-day operations, while NIH program staff would have substantial involvement in coordinating milestones, standards, expectations for sharing data and resources, and integration with other components of the broader initiative.

How much funding could an applicant request?

The FOA allowed applicants to request up to $12.4 million per year in total costs, for project periods of up to five years.

What was the estimated total program funding level?

The initiative was described as having an estimated total program funding level of roughly $62 million across the life of the program.

How many awards did NIH expect to make?

NIH anticipated making between one and three awards, depending on the scientific and operational merit of applications and the availability of funds.

Was cost sharing or matching required?

No. The FOA states there was no cost sharing or matching requirement.

Who was eligible to apply?

Eligibility was broad and included domestic institutions and organizations capable of operating sophisticated mouse production and cryopreservation programs at scale. Eligible applicants included small businesses, for-profit entities (including those other than small businesses), nonprofit organizations with or without 501(c)(3) status, private and public institutions of higher education, and state governments. The FOA also noted eligibility for institution types such as HBCUs, Hispanic-serving Institutions, TCCUs, Alaska Native and Native Hawaiian Serving Institutions, and eligible federal agencies.

Which NIH organization issued this opportunity?

The sponsoring agency was the National Institutes of Health, and the opportunity was issued through the Office of Strategic Coordination, reflecting NIH-wide and cross-institute relevance.

When was the opportunity posted and when did it close?

The opportunity was posted on September 10, 2010. The original and current closing date listed was November 10, 2010.

When was the opportunity archived?

The FOA was archived on December 11, 2010.

What role does the KOMP Repository play in this program?

The KOMP Repository is part of the long-term preservation and distribution strategy described in the FOA. Awardees were expected to cryopreserve germplasm from produced mouse lines and send aliquots of those cryopreserved materials to the KOMP Repository to ensure long-term preservation, redundancy, and distribution capability.

What is the overall pipeline this FOA supports?

The FOA supports a pipeline that starts with engineered, gene-targeted ES cell lines from international repositories and ends with standardized phenotype data. The U42-funded production and cryopreservation sites serve as the operational middle of this pipeline by generating validated knockout mice, shipping them to phenotyping centers, and banking germplasm for long-term access.

What broader scientific goal is this program trying to achieve?

The overarching objective is to enable systematic understanding of gene function by supporting the production and phenotyping of knockout mice for essentially every protein-coding gene in the mammalian genome, creating reliable and renewable resources that can be used across the biomedical community to study gene function, disease mechanisms, and potential therapeutic targets.

Why does the FOA reference many CFDA program numbers across NIH mission areas?

The FOA spans many NIH mission areas because knockout mouse resources are foundational tools used across a wide range of biomedical research fields (including, as noted, areas such as genome research, cancer biology, cardiovascular research, neuroscience, mental health, aging, and child health). The broad CFDA coverage reflects the cross-cutting utility of these models.