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Strain
C57Bl/6J-Lyplal1em1Espel/Mmmh

Summary Data Summary
Official Name C57Bl/6J-Lyplal1em1Espel/Mmmh
Common Name C57Bl/6J-Lyplal1em1Espel/Mmmh
Description CRISPR/Cas9 technology was used by the University of
Michigan Transgenic Animal Model Core to generate a
genetically modified C57BL6/J mouse strain with a one base
pair deletion in the first exon of Lyplal1 referred to as
Lyplal1 KO mice. Five single guide RNA (sgRNA) targets were
identified for testing in exon 1 Ensembl.org exon
id=ENSMUSE00000375205 with the algorithm described by Hsu
and colleagues (PMID:23873081) and cloned into plasmid pX330
(Addgene plasmid #42230), as described (PMID:24157548). One
sgRNA and protospacer adjacent motif (PAM) with the highest
chromosome cleavage activity and a high specificity
prediction was selected for generation of Lyplal1 KO mice:
5’ GGGACACCACACAACGCGGC 3’ PAM: AGG. Mouse zygote
microinjection was carried out as described
(DOI:10.1007/978-3-642-20792-1_6). Animals were housed in an
AAALAC accredited facility in accordance with the National
Research Council’s guide for the care and use of laboratory
animals. Procedures were approved by the University of
Michigan’s Institutional Animal Care & Use Committee. pX330
plasmid DNA expressing the active sgRNA was purified with an
endotoxin-free kit. Plasmid DNA concentration was adjusted
to 5 ng/ul for pronuclear microinjection. Mouse zygotes for
microinjection were obtained by mating superovulated
C57BL/6J females with males of the same strain (JAX Strain
#000664). Mouse zygotes (457) were microinjected and those
that survived injection (395) were transferred to
pseudopregnant females. Genomic DNA was isolated from tail
tip biopsies of 65 potential founders that were born and
analyzed for CRISPR/Cas9 induced indels with CEL I
endonuclease. A total of 26 G0 mouse pups (40%) were
identified as carrying mutations in Lyplal1. The efficiency
of the producing mutant mouse founders exceeded the
efficiency of producing transgenic mice carrying random
integrations of DNA transgenes by a factor of four.
Development Status Phenotyping ongoing
Creation Method knockout
TypeCount
Investigators 1
Genomics - Modifications 1
Experiments 2


Investigators
NameInstitution
Brian HalliganUniversity of Michigan-Ann Arbor


Genomic Information
GeneAllele 1Allele 2Protocol
Lyplal1knockoutknockoutNot Specified


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