ABSTRACT
JHU researchers generated mice carrying a targeted deletion of recently characterized Pac gene. CRISPR/Cas9-mediated homologous recombination was performed to generate Pac floxed allele in mice. ES cell clones with proper targeting were identified by PCR and sequencing analysis. Two correctly targeted ES clones were chosen to generate chimeric mice by blastocyst injection, which produced germline-transmitted offspring. Mice with Pac floxed allele were mated with EIIa-Cre (JAX #3724) to generate Pac+/− mice. Cre-mediated recombination deleted exon 2, resulting in frameshift mutation before TM1 domain and creating a null allele. Intercrossing of the heterozygous knockouts yielded viable and fertile homozygous Pac knockout mice.
FEATURES
JHU researcher characterized novel gene called proton-associated channel Pac, previously known as TMEM206. PAC is unlike any previously identified ion channel. This gene is evolutionarily conserved and is essential in proton-associated Cl- currents. JHU researchers show that PAC contributes to acid-induced cell death and ischemic brain injury. Pac KO mice could be used to further explore the physiological role of PAC in hypoxic conditions and regulation of proton-associated Cl- currents.
STAGE OF DEVELOPMENT
The animal was used to study PAC role in regulating proton-associated Cl- currents ischemic brain injury in vivo.
ASSOCIATED PUBLICATIONS
Science 26 Apr 2019: Vol. 364, Issue 6438, pp. 395-399
