Lentiviral vector mediated gene therapy for type I Dent disease ameliorates Dent disease-like phenotypes for three months in ClC-5 null mice.

Type 1 Dent disease is caused by changes in chloride voltage-gated channel 5 () gene on chromosome X, which causes the lack or dysfunction of chloride channel ClC-5. Affected subjects show proteinuria and hypercalciuria, and eventually develop end-stage kidney disease. Currently there is no cure for this disease.

Targeting DNA polymerase to DNA double-strand breaks reduces DNA deletion size and increases templated insertions generated by CRISPR/Cas9.

Most insertions or deletions generated by CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9) endonucleases are short (<25 bp), but unpredictable on-target long DNA deletions (>500 bp) can be observed. The possibility of generating long on-target DNA deletions poses safety risks to somatic genome editing and makes the outcomes of genome editing less predictable. Methods for generating refined mutations are desirable but currently unavailable.

Adenine Base Editor Ribonucleoproteins Delivered by Lentivirus-Like Particles Show High On-Target Base Editing and Undetectable RNA Off-Target Activities.

Adenine base editors (ABEs) can correct gene mutations without creating double-strand breaks. However, in recent reports, these editors showed guide-independent RNA off-target activities. This work describes our development of a delivery method to minimize ABEs' RNA off-target activity.

Sensitive and reliable evaluation of single-cut sgRNAs to restore dystrophin by a GFP-reporter assay.

Most Duchenne muscular dystrophy (DMD) cases are caused by deletions or duplications of one or more exons that disrupt the reading frame of DMD mRNA. Restoring the reading frame allows the production of partially functional dystrophin proteins, and result in less severe symptoms. Antisense oligonucleotide mediated exon skipping has been approved for DMD, but this strategy needs repeated treatment.

A tissue-engineered uterus supports live births in rabbits.

Bioengineered uterine tissue could provide a treatment option for women with uterine factor infertility. In large animal models, reconstruction of the uterus has been demonstrated only with xenogeneic tissue grafts. Here we use biodegradable polymer scaffolds seeded with autologous cells to restore uterine structure and function in rabbits.

CRISPR/Cas9 increases mitotic gene conversion in human cells.

Gene conversion is a process of transferring genetic material from one homologous sequence to another. Most reported gene conversions are meiotic although mitotic gene conversion is also described. When using CRISPR/Cas9 to target the human hemoglobin subunit beta (HBB) gene, hemoglobin subunit delta (HBD) gene footprints were observed in HBB gene.

Delivering SaCas9 mRNA by lentivirus-like bionanoparticles for transient expression and efficient genome editing.

The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) system discovered using bacteria has been repurposed for genome editing in human cells. Transient expression of the editor proteins (e.g.

Large-Scale Preparation of Extracellular Vesicles Enriched with Specific microRNA.

This article describes a method for producing microRNA (miRNA)-enriched extracellular vesicles in large quantities. It enables in vivo delivery of specific miRNA for therapeutic applications.

Characterization of Barium Borate Frameworks Using Raman Spectroscopy.

Systematic micro-Raman scattering investigations have been carried out on Sm doped 2(BaO)-n(BO) matrices for n = 4, 5, 8, and 2(BaO)-(NaO)-9(BO) using the 364 nm excitation of an Ar laser. The Raman results have been compared with the known structures of barium tetraborate, barium pentaborate, barium octaborate, and barium sodium nonaborate. An excellent correlation has been found between the BO/BO composition ratios for each product and intensity ratios of the designated BO and BO Raman peaks.

Prediction of enophthalmos by computer-based volume measurement of orbital fractures in a Korean population.

Purpose: To estimate posttraumatic enophthalmos using computer-based volume measurement of the orbital fracture to provide information on surgical guidelines.

Methods: The fracture volume of orbital wall fractures in 35 patients who did not undergo surgery was measured using a Rapidia work station system. Hertel ophthalmometry, diplopia, and ocular motility were investigated.

Amniotic membrane corrects surgically induced astigmatism.

Human amniotic membrane (AM) has been widely used for the reconstruction of ocular surface. We conducted this study to investigate whether AM shows efficacy in maintaining the correction of astigmatism or not. We performed photoastigmatic refractive keratectomy (PARK) on rabbits and grafted AM on the cornea.

Application of newly developed amniotic membrane ointment for photorefractive keratectomy in rabbits.

We developed amniotic membrane ointment (AMO), and the effect of instilling the AMO after photorefractive keratectomy (PRK) was investigated with respect to inflammatory cell infiltration into the corneal stroma, apoptosis of keratocytes, and suppression of lipid peroxidation of cellular walls. The PRK procedure was performed on both eyes of 10 white rabbits. One eye of each rabbit (the experimental eye) was instilled with the AMO and the other eye of the rabbit (the control eye) with a base ointment 0, 8 and 16 h after the PRK procedure.

Early mitochondrial hyperpolarization and intracellular alkalinization in lactacystin-induced apoptosis of retinal pigment epithelial cells.

We investigated the induction and underlying mechanism of apoptosis in retinal pigment epithelial cells by the inhibition of proteasome activity using lactacystin. Rat retinal pigment epithelial cell line retinal pigment epithelial (RPE)-J was used in this study. Apoptosis was evaluated by light and electron microscopies, DNA electrophoresis, and terminal deoxynucleotidyl transferase dUTP nick-end labeling assay.

Diminished hepatic expression of the HNF-6 transcription factor during bile duct obstruction.

Hepatocyte nuclear factor 6 (HNF-6) is a member of the one cut family of transcription factors and potentially regulates expression of numerous target genes important for hepatocyte function. In the liver, HNF-6 is expressed not only in hepatocytes, but also in biliary epithelial cells (BEC). To evaluate the in vivo function of HNF-6, we examined the hepatic expression pattern of HNF-6 messenger RNA (mRNA) and protein after bile duct ligation (BDL)-mediated liver injury.