Our lab focuses on the developmental pathways and factors that are critical for building the cardiopulmonary system. Using a combination of mouse genetics, biochemistry, and genomic analysis, we seek to better understand how the lung and heart develop, how developmental pathways are disrupted in human cardiopulmonary disease, and whether such pathways and factors can be harnessed to promote pulmonary and cardiac regeneration in the adult.
"Exons 12, 13, and 14, that encode the forkhead DNA-binding domain, were replaced with a Neo cassette via homologous recombination. RT-PCR and immunohistochemistry confirmed absence of mRNA in embryos and protein in heart tissue."
"A loxP site was inserted upstream of exon 10 and an FRT flanked neo cassette and second loxP site were inserted downstream of exon 13 via homologous recombination. Flp mediated recombination removed the neo cassette."
"A targeting vector was designed to replace the majority of the forkhead domain (exons 12 and 13) with a neomycin resistance gene. Immunohistochemistry of mutant embryos confirmed lack of expression."
"A loxP site was inserted upstream of exon 12. An FRT flanked neo cassette with a 3' loxP site was inserted downstream of exon 13. Flp mediated recombination removed the neo cassette and left exons 12 and 13 floxed."
"The single coding exon was flanked by loxP sites and followed by neomycin cassette that was subsequently removed via flp recombination"
"This mutant allele retains the 5' region of the gene through the first 20 base pairs of exon 2, which encode the first zinc finger of the protein. A lacZ-PGK-neo cassette has replaced the remaining 3' portion of exon2 and all of intron 2 and exon3, including the second and third zinc finger coding sequences. Neither RT-qPCR analysis nor in situ hybridization to E10.5 embryos detected any transcripts."
"A null allele in mice was generated by replacing all of the coding exons with the bacterial lacZ gene."
"An FRT-flanked neo cassette with a 3' loxP site was inserted upstream of exon 2. An additional loxP site was inserted downstream of exon 2. Flp-mediated recombination removed the neo cassette."
"The miR302-367flox/flox allele was generated by flanking all five of the miRNAs in this cluster with loxP sites using standard homologous recombination in ES cells."
"The R26R-miR302-367Tg/+ allele was generated using previously described vectors to insert a DNA sequence containing all five members of the miR302-367 cluster into the CAG-R26R locus using standard homologous recombination in ES cells (PMID: 15093544, PMID: 20382891)."
"The human promoter drives lung-specifc expression of the mouse coding region. Three founders were generated. The pound symbol (#) is used when no line is specified and/or lines are pooled."
Mice carrying the Fzd2flox/flox allele were crossed into the Shhcre allele to delete Fzd2 specifically in the lung epithelium.
"The coding region in exon 1 was replaced with a reverse tet-activator (rtTA). A downstream FRT flanked neo cassette was removed by germ line, flp mediated recombination."
"Germ line, cre mediated recombination removed exon 2 and 3."
"A loxP site was inserted upstream of exon 2 and an frt flanked neo cassette with a 5' loxP site was inserted downstream of exon 3."
"A targeting construct was designed so that homologous recombination in ES cells inserted the cre/ERT2 fusion sequence and a neo selection marker after the ATG start codon in exon 1 (replacing exon 1). After establishing germline transmission, Flp-mediated excision removed the neo cassette."
"A targeting vector was designed to replace exon 1, including the ATG, with a lacZ-neo cassette. Unexpectedly, transcripts contining exons 3 and 4 were detected at low levels. LacZ expression recapitulates the endogenous expression pattern."