@article {2042, title = {Hardwiring tissue-specific AAV transduction in mice through engineered receptor expression.}, journal = {Nat Methods}, volume = {20}, year = {2023}, month = {2023 Jul}, pages = {1070-1081}, abstract = {

The development of transgenic mouse models that express genes of interest in specific cell types has transformed our understanding of basic biology and disease. However, generating these models is time- and resource-intensive. Here we describe a model system, SELective Expression and Controlled Transduction In Vivo (SELECTIV), that enables efficient and specific expression of transgenes by coupling adeno-associated virus (AAV) vectors with Cre-inducible overexpression of the multi-serotype AAV receptor, AAVR. We demonstrate that transgenic AAVR overexpression greatly increases the efficiency of transduction of many diverse cell types, including muscle stem cells, which are normally refractory to AAV transduction. Superior specificity is achieved by combining Cre-mediated AAVR overexpression with whole-body knockout of endogenous Aavr, which is demonstrated in heart cardiomyocytes, liver hepatocytes and cholinergic neurons. The enhanced efficacy and exquisite specificity of SELECTIV has broad utility in development of new mouse model systems and expands the use of AAV for gene delivery in vivo.

}, keywords = {Animals, Dependovirus, Gene Transfer Techniques, Genetic Therapy, Genetic Vectors, Mice, Mice, Transgenic, Transduction, Genetic, Transgenes}, issn = {1548-7105}, doi = {10.1038/s41592-023-01896-x}, author = {Zengel, James and Wang, Yu Xin and Seo, Jai Woong and Ning, Ke and Hamilton, James N and Wu, Bo and Raie, Marina and Holbrook, Colin and Su, Shiqi and Clements, Derek R and Pillay, Sirika and Puschnik, Andreas S and Winslow, Monte M and Idoyaga, Juliana and Nagamine, Claude M and Sun, Yang and Mahajan, Vinit B and Ferrara, Katherine W and Blau, Helen M and Carette, Jan E} } @article {978, title = {CRISPR GENOME SURGERY IN THE RETINA IN LIGHT OF OFF-TARGETING.}, journal = {Retina}, volume = {38}, year = {2018}, month = {2018 08}, pages = {1443-1455}, abstract = {

PURPOSE: Recent concerns regarding the clinical utilization of clustered regularly interspaced short palindromic repeats (CRISPR) involve uncertainties about the potential detrimental effects that many arise due to unintended genetic changes, as in off-target mutagenesis, during CRISPR genome surgery. This review gives an overview of off-targeting detection methods and CRISPR{\textquoteright}s place in the clinical setting, specifically in the field of ophthalmology.

RESULTS: As CRISPR utilization in the laboratory setting has increased, knowledge regarding CRISPR mechanisms including its off-target effects has also increased. Although a perfect method for achieving 100\% specificity is yet to be determined, the past few years have seen many developments in off-targeting detection and in increasing efficacy of CRISPR tools.

CONCLUSION: The CRISPR system has high potential to be an invaluable therapeutic tool as it has the ability to modify and repair pathogenic retinal lesions. Although it is not yet a perfect system, with further efforts to improve its specificity and efficacy along with careful screening of off-target mutations, CRISPR-mediated genome surgery potential can become maximized and applied to patients.

}, keywords = {Clustered Regularly Interspaced Short Palindromic Repeats, Gene Editing, Genetic Therapy, Humans, Ophthalmology, Retinal Diseases}, issn = {1539-2864}, doi = {10.1097/IAE.0000000000002197}, author = {Cho, Galaxy Y and Schaefer, Kellie A and Bassuk, Alexander G and Tsang, Stephen H and Mahajan, Vinit B} } @article {637, title = {Reprogramming metabolism by targeting sirtuin 6 attenuates retinal degeneration.}, journal = {J Clin Invest}, volume = {126}, year = {2016}, month = {2016 Dec 01}, pages = {4659-4673}, abstract = {

Retinitis pigmentosa (RP) encompasses a diverse group of Mendelian disorders leading to progressive degeneration of rods and then cones. For reasons that remain unclear, diseased RP photoreceptors begin to deteriorate, eventually leading to cell death and, consequently, loss of vision. Here, we have hypothesized that RP associated with mutations in phosphodiesterase-6 (PDE6) provokes a metabolic aberration in rod cells that promotes the pathological consequences of elevated cGMP and Ca2+, which are induced by the Pde6 mutation. Inhibition of sirtuin 6 (SIRT6), a histone deacetylase repressor of glycolytic flux, reprogrammed rods into perpetual glycolysis, thereby driving the accumulation of biosynthetic intermediates, improving outer segment (OS) length, enhancing photoreceptor survival, and preserving vision. In mouse retinae lacking Sirt6, effectors of glycolytic flux were dramatically increased, leading to upregulation of key intermediates in glycolysis, TCA cycle, and glutaminolysis. Both transgenic and AAV2/8 gene therapy-mediated ablation of Sirt6 in rods provided electrophysiological and anatomic rescue of both rod and cone photoreceptors in a preclinical model of RP. Due to the extensive network of downstream effectors of Sirt6, this study motivates further research into the role that these pathways play in retinal degeneration. Because reprogramming metabolism by enhancing glycolysis is not gene specific, this strategy may be applicable to a wide range of neurodegenerative disorders.

}, keywords = {Animals, Cellular Reprogramming, Cellular Reprogramming Techniques, Citric Acid Cycle, Cyclic GMP, Cyclic Nucleotide Phosphodiesterases, Type 6, Dependovirus, Disease Models, Animal, Eye Proteins, Genetic Therapy, Glycolysis, Mice, Mice, Mutant Strains, Mutation, Retinal Rod Photoreceptor Cells, Retinitis Pigmentosa, Sirtuins, Transduction, Genetic}, issn = {1558-8238}, doi = {10.1172/JCI86905}, author = {Zhang, Lijuan and Du, Jianhai and Justus, Sally and Hsu, Chun-Wei and Bonet-Ponce, Luis and Wu, Wen-Hsuan and Tsai, Yi-Ting and Wu, Wei-Pu and Jia, Yading and Duong, Jimmy K and Mahajan, Vinit B and Lin, Chyuan-Sheng and Wang, Shuang and Hurley, James B and Tsang, Stephen H} }