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128 changes: 128 additions & 0 deletions joss.06474/10.21105.joss.06474.crossref.xml
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<full_title>Journal of Open Source Software</full_title>
<abbrev_title>JOSS</abbrev_title>
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<title>MousebreedeR: A novel software to assist in the design
of breeding schema for complex genotypes of experimental
organisms</title>
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<contributors>
<person_name sequence="first" contributor_role="author">
<given_name>Mike</given_name>
<surname>Sportiello</surname>
<ORCID>https://orcid.org/0000-0003-1690-8702</ORCID>
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contributor_role="author">
<given_name>David J.</given_name>
<surname>Topham</surname>
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<citation key="RN204">
<article_title>Simplified method to measure mouse
fertility</article_title>
<author>Handelsman</author>
<journal_title>Endocrinology</journal_title>
<issue>8</issue>
<volume>161</volume>
<doi>10.1210/endocr/bqaa114</doi>
<issn>1945-7170</issn>
<cYear>2020</cYear>
<unstructured_citation>Handelsman, D. J., Walters, K. A.,
&amp; Ly, L. P. (2020). Simplified method to measure mouse fertility.
Endocrinology, 161(8).
https://doi.org/10.1210/endocr/bqaa114</unstructured_citation>
</citation>
<citation key="RN203">
<article_title>Mouse gestation length is genetically
determined</article_title>
<author>Murray</author>
<journal_title>PLoS One</journal_title>
<issue>8</issue>
<volume>5</volume>
<doi>10.1371/journal.pone.0012418</doi>
<issn>1932-6203</issn>
<cYear>2010</cYear>
<unstructured_citation>Murray, S. A., Morgan, J. L., Kane,
C., Sharma, Y., Heffner, C. S., Lake, J., &amp; Donahue, L. R. (2010).
Mouse gestation length is genetically determined. PLoS One, 5(8),
e12418.
https://doi.org/10.1371/journal.pone.0012418</unstructured_citation>
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207 changes: 207 additions & 0 deletions joss.06474/paper.jats/10.21105.joss.06474.jats
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<!DOCTYPE article PUBLIC "-//NLM//DTD JATS (Z39.96) Journal Publishing DTD v1.2 20190208//EN"
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<journal-meta>
<journal-id></journal-id>
<journal-title-group>
<journal-title>Journal of Open Source Software</journal-title>
<abbrev-journal-title>JOSS</abbrev-journal-title>
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<issn publication-format="electronic">2475-9066</issn>
<publisher>
<publisher-name>Open Journals</publisher-name>
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<article-meta>
<article-id pub-id-type="publisher-id">6474</article-id>
<article-id pub-id-type="doi">10.21105/joss.06474</article-id>
<title-group>
<article-title>MousebreedeR: A novel software to assist in the design of
breeding schema for complex genotypes of experimental
organisms</article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author" corresp="yes">
<contrib-id contrib-id-type="orcid">https://orcid.org/0000-0003-1690-8702</contrib-id>
<name>
<surname>Sportiello</surname>
<given-names>Mike</given-names>
</name>
<xref ref-type="aff" rid="aff-1"/>
<xref ref-type="aff" rid="aff-2"/>
<xref ref-type="corresp" rid="cor-1"><sup>*</sup></xref>
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<name>
<surname>Topham</surname>
<given-names>David J.</given-names>
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<aff id="aff-1">
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<institution>Center for Vaccine Biology and Immunology, University of
Rochester Medical Center, Rochester, NY 14642, United States of
America</institution>
</institution-wrap>
</aff>
<aff id="aff-2">
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<institution>Medical Scientist Training Program, University of Rochester
Medical Center, Rochester, NY 14642, United States of
America</institution>
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<corresp id="cor-1">* E-mail: <email></email></corresp>
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<day>28</day>
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<year>2023</year>
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<kwd-group kwd-group-type="author">
<kwd>breeding</kwd>
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<sec id="summary">
<title>Summary</title>
<p>With the advent of gene editing playing a regular role in producing
new experimental models, the need to get the appropriate allele at
each respective locus in a model organism is now common. Instead of
simple “knockout” vs “wildtype” mouse experiments, for example, it is
common for there to be a gene of interest that is either floxed
(surrounded by known genetic sequence that allow for a gene to be
knocked out when coexpression of the cre protein is expressed) or not
floxed depending on the experimental group, another gene that marks
cells of a certain phenotype such as a fluorescent reporter, a cre
expression cassette to flox the gene of interest out, and more. While
it is often obvious how to efficiently breed experimental organisms to
obtain one locus of interest, once three or more loci are involved
this becomes difficult to assess. MousebreedeR is a free and open
source web-based shiny app and command line R package that takes
desired mouse genotypes as well as the genotypes of potential mouse
breeders that the user already has on hand as inputs, and delivers
efficient breeding schema as the output. In addition, it supplies the
user with the probability of producing each potential pup, the
probability that no pups with the desired genotype are born in a
litter, and more. We have also supplied code that assists the user in
plotting these tables to help them visualize crosses of interest and
make decisions in their breeding.</p>
</sec>
<sec id="statement-of-need">
<title>Statement of need</title>
<p>Efficient breeding is a scientific imperative: with a limited
period of fertility in experimental organisms, users may only have
limited attempts with organisms of desired genotypes
(<xref alt="Handelsman et al., 2020" rid="ref-RN204" ref-type="bibr">Handelsman
et al., 2020</xref>;
<xref alt="Murray et al., 2010" rid="ref-RN203" ref-type="bibr">Murray
et al., 2010</xref>). Taking into account both gestational time and
the necessary time it takes to wean pups off their mother’s milk, an
inefficient cross can set the user back months in the case of a female
pregnant with the non-optimal male’s offspring. Furthermore, as most
research centers pay <italic>per diem</italic> costs, inefficiency is
a large direct cost for many experimentalists. Individual cages often
cost hundreds of dollars per year just in maintenance fees, not even
accounting for the cost of extra rounds of genotyping (requiring PCR
primers, PCR mastermix, gels, dye, etc) or for the time dedicated to
the project by employees. Most importantly, ethical and animal welfare
concerns also arise as inefficient breeding results in the mass
euthanasia of non-desired animals as well as ear/tail-clipping
required for genotyping, which may cause animals pain and
distress.</p>
<p>While the breeding schema to obtain a litter of full knockout mice
from one wildtype/wildtype parent and one knockout/knockout parent is
straightforward, this is not the case for the situation where one has
4 alleles at 4 loci in 4 separate mice, which all need to co-occur in
the same mouse for a given planned experiment, for example.
Furthermore, to our knowledge, no current software exists that can
quantitatively assist the user in creating their breeding schema.
Indeed, if there are 2 alleles at each locus, when attempting to make
a genetically marked, inducible, cre-lox model with T cell specificity
as our lab was doing and which prompted us to create this software, 81
possible combinations exist (3 (AA, Aa, and aa) to the 4th power).
With 81 possible males and 81 possible females to mate, 6561 possible
pairings exist (81 * 81). Use of the mousebreedeR software to optimize
that breeding schema has been most helpful.</p>
</sec>
<sec id="acknowledgements">
<title>Acknowledgements</title>
<p>This work was supported by the National Institutes of Health (NIH)
National Institute of Allergy and Infectious Diseases Grant
P01-AI102851-10. M.S. was supported by the NIH/National Heart, Lung,
and Blood Institute Training Grant T32-HL066988-20, as well as by the
NIH Training Grant/National Institute of General Medical Sciences
Grant T32GM007356-48.</p>
</sec>
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<person-group person-group-type="author">
<name><surname>Handelsman</surname><given-names>D. J.</given-names></name>
<name><surname>Walters</surname><given-names>K. A.</given-names></name>
<name><surname>Ly</surname><given-names>L. P.</given-names></name>
</person-group>
<article-title>Simplified method to measure mouse fertility</article-title>
<source>Endocrinology</source>
<year iso-8601-date="2020">2020</year>
<volume>161</volume>
<issue>8</issue>
<issn>1945-7170</issn>
<uri>https://www.ncbi.nlm.nih.gov/pubmed/32645712</uri>
<pub-id pub-id-type="doi">10.1210/endocr/bqaa114</pub-id>
</element-citation>
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<ref id="ref-RN203">
<element-citation publication-type="article-journal">
<person-group person-group-type="author">
<name><surname>Murray</surname><given-names>S. A.</given-names></name>
<name><surname>Morgan</surname><given-names>J. L.</given-names></name>
<name><surname>Kane</surname><given-names>C.</given-names></name>
<name><surname>Sharma</surname><given-names>Y.</given-names></name>
<name><surname>Heffner</surname><given-names>C. S.</given-names></name>
<name><surname>Lake</surname><given-names>J.</given-names></name>
<name><surname>Donahue</surname><given-names>L. R.</given-names></name>
</person-group>
<article-title>Mouse gestation length is genetically determined</article-title>
<source>PLoS One</source>
<year iso-8601-date="2010">2010</year>
<volume>5</volume>
<issue>8</issue>
<issn>1932-6203</issn>
<uri>https://www.ncbi.nlm.nih.gov/pubmed/20811634</uri>
<pub-id pub-id-type="doi">10.1371/journal.pone.0012418</pub-id>
<fpage>e12418</fpage>
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