Simultaneous deletion of floxed genes mediated by CaMKIIa-Cre in the brain and in male germ cells: application to conditional and conventional disruption of Go-alfa

The Cre/LoxP system is a well-established approach to spatially and temporally control genetic inactivation. The calcium/calmodulin-dependent protein kinase II alpha subunit (CaMKIIα) promoter limits expression to specific regions of the forebrain and thus has been utilized for the brain-specific inactivation of the genes. Here, we show that CaMKIIα-Cre can be utilized for simultaneous inactivation of genes in the adult brain and in male germ cells. Double transgenic Rosa26+/stop-lacZ::CaMKIIα-Cre+/Cre mice generated by crossing CaMKIIα-Cre+/Cre mice with floxed ROSA26 lacZ reporter (Rosa26+/stop-lacZ) mice exhibited lacZ expression in the brain and testis. When these mice were mated to wild-type females, about 27% of the offspring were whole body blue by X-gal staining without inheriting the Cre transgene. These results indicate that recombination can occur in the germ cells of male Rosa26+/stop-lacZ::CaMKIIα-Cre+/Cre mice. Similarly, when double transgenic Gnao+/f::CaMKIIα-Cre+/Cre mice carrying a floxed Go-alpha gene (Gnaof/f) were backcrossed to wild-type females, approximately 22% of the offspring carried the disrupted allele (GnaoΔ) without inheriting the Cre transgene. The GnaoΔ/Δ mice closely resembled conventional Go-alpha knockout mice (Gnao−/−) with respect to impairment of their behavior. Thus, we conclude that CaMKIIα-Cre mice afford recombination for both tissue- and time-controlled inactivation of floxed target genes in the brain and for their permanent disruption. This work also emphasizes that extra caution should be exercised in utilizing CaMKIIα-Cre mice as breeding pairs.Fil: Choi, Chan-Il. Ajou University. School of Medicine; Corea del SurFil: Yoon, Sang-Phil. Ajou University. School of Medicine; Corea del SurFil: Choi, Jung-Mi. Ajou University. School of Medicine; Corea del SurFil: Kim, Sung-Soo. Ajou University. School of Medicine; Corea del SurFil: Lee, Young-Don. Ajou University. School of Medicine; Corea del SurFil: Birnbaumer, Lutz. National Institute of Environmental Health Sciences; Estados Unidos. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; ArgentinaFil: Suh-Kim. Haeyoung. Ajou University. School of Medicine; Corea del Su

Two novel mutations of Wiskott–Aldrich syndrome: the molecular prediction of interaction between the mutated WASP L101P with WASP-interacting protein by molecular modeling

AbstractWiskott–Aldrich syndrome (WAS) is an X-linked disorder characterized by eczema, thrombocytopenia and increased susceptibility of infections, with mutations of the WAS gene being responsible for WAS and X-linked thrombocytopenia. Herein, two novel mutations of WAS at T336C on exon 3, and at 1326–1329, a G deletion on exon 10, resulting in L101P missense mutation and frameshift mutation 444 stop, respectively, are reported. The affected patients with either mutation showed severe suppression of WAS protein (WASP) levels, T cell proliferation, and CFSE-labeled T cells division. Because WASP L101 have not shown direct nuclear Overhauser effect (NOE) contact with the WASP-interacting protein (WIP) in NMR spectroscopy, molecular modeling was performed to evaluate the molecular effect of WASP P101 to WIP peptide. It is presumed that P101 induced a conformational change in the Q99 residue of WASP and made the side chain of Q99 move away from the WIP peptide, resulting in disruption of the hydrogen bond between Q99 WASP and Y475 WIP. A possible model for the molecular pathogenesis of WAS has been proposed by analyzing the interactions of WASP and WIP using a molecular modeling study

Investigation of the SH3BP2 Gene Mutation in Cherubism

Cherubism is a rare developmental lesion of the jaw that is generally inherited as an autosomal dominant trait. Recent studies have revealed point mutations in the SH3BP2 gene in cherubism patients. In this study, we examined a 6-year-old Korean boy and his family. We found a Pro418Arg mutation in the SH3BP2 gene of the patient and his mother. A father and his 30-month-old younger brother had no mutations. Immunohistochemically, the multinucleated giant cells proved positive for CD68 and tartrate-resistant acid phosphatase (TRAP). Numerous spindle-shaped stromal cells expressed a ligand for receptor activator of nuclear factor kB (RANKL), but not in multinucleated giant cells. These results provide evidence that RANKL plays a critical role in the differentiation of osteoclast precursor cells to multinucleated giant cells in cherubism. Additionally, genetic analysis may be a useful method for differentiation of cherubism.</p

Comb-rooted multi-channel synthesis of ultra-narrow optical frequencies of few Hz linewidth

We report a multi-channel optical frequency synthesizer developed to generate extremely stable continuous wave lasers directly out of the optical comb of an Er-doped fiber oscillator. Being stabilized to a high-finesse cavity with a fractional frequency stability of $3.8\times10^{-15}$ at 0.1 s, the comb-rooted synthesizer produces multiple optical frequencies of ultra-narrow linewidth of 1.0 Hz at 1 s concurrently with an output power of tens of mW per each channel. Diode-based stimulated emission by injection locking is a key mechanism that allows comb frequency modes to sprout up with sufficient power amplification but no loss of original comb frequency stability. Channel frequencies are individually selectable with a 0.1 GHz increment over the entire comb bandwidth spanning 4.25 THz around a 1550 nm center wavelength. A series of out-of-loop test results is discussed to demonstrate that the synthesizer is able to provide stable optical frequencies with the potential for advancing diverse ultra-precision applications such as optical clocks comparison, atomic line spectroscopy, photonic microwaves generation, and coherent optical telecommunications.Comment: 19 pages, 4 figure

EM-Network: Oracle Guided Self-distillation for Sequence Learning

We introduce EM-Network, a novel self-distillation approach that effectively leverages target information for supervised sequence-to-sequence (seq2seq) learning. In contrast to conventional methods, it is trained with oracle guidance, which is derived from the target sequence. Since the oracle guidance compactly represents the target-side context that can assist the sequence model in solving the task, the EM-Network achieves a better prediction compared to using only the source input. To allow the sequence model to inherit the promising capability of the EM-Network, we propose a new self-distillation strategy, where the original sequence model can benefit from the knowledge of the EM-Network in a one-stage manner. We conduct comprehensive experiments on two types of seq2seq models: connectionist temporal classification (CTC) for speech recognition and attention-based encoder-decoder (AED) for machine translation. Experimental results demonstrate that the EM-Network significantly advances the current state-of-the-art approaches, improving over the best prior work on speech recognition and establishing state-of-the-art performance on WMT'14 and IWSLT'14.Comment: ICML 202