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Human OPRM1 and murine Oprm1 promoter driven viral constructs for genetic access to μ-opioidergic cell types
Tools for gaining long-term genetic access to mu-opioid receptor (MOR) neural cell types are limited. Here, the authors develop a suite of adeno-associated viral tools allowing selective genetic access to MOR cell types, and showcase their use across species.
- Gregory J. Salimando
- , Sébastien Tremblay
- & Gregory Corder
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Article
| Open Access
A mosaic adeno-associated virus vector as a versatile tool that exhibits high levels of transgene expression and neuron specificity in primate brain
Viral vectors with intense and long-lasting transgene expression are essential for manipulating and imaging of neuronal activity in the primate brain. Here, the authors created a mosaic vector composed of capsid proteins derived from AAV1 and AAV2 that exhibits improved transgene expression and neuronal specificity.
- Kei Kimura
- , Yuji Nagai
- & Masahiko Takada
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| Open Access
Functional gene delivery to and across brain vasculature of systemic AAVs with endothelial-specific tropism in rodents and broad tropism in primates
Delivering genes to and across the brain vasculature efficiently and specifically across species remains challenging. Here, the authors show that endothelial-specific AAVs with serotype flexibility enable redosing and transform the brain vasculature into an in vivo biofactory in genetically diverse rodents. In primates, these vectors cross the blood-brain-barrier and show broad tropism.
- Xinhong Chen
- , Damien A. Wolfe
- & Viviana Gradinaru
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| Open Access
The AAV capsid can influence the epigenetic marking of rAAV delivered episomal genomes in a species dependent manner
rAAV vectors vary in their effectiveness between species, making it difficult to predict clinical outcomes. Here authors show that AAV capsid proteins influence the vector epigenomic state in cells, and that a single amino acid change in the vector can alter the vector epigenome and hence transgene expression levels between species.
- Adriana Gonzalez-Sandoval
- , Katja Pekrun
- & Mark A. Kay
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Article
| Open Access
Cross-species evolution of a highly potent AAV variant for therapeutic gene transfer and genome editing
Gonzalez et al. use a viral evolutionary approach to generate cross-species compatible AAV (ccAAVs) vectors. They describe a highly potent new variant, AAV.cc47, with enhanced transduction efficiency over AAV serotype 9 and show its efficacy in different mouse models, pigs and non-human primates.
- Trevor J. Gonzalez
- , Katherine E. Simon
- & Aravind Asokan
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Article
| Open Access
Cell-specific regulation of gene expression using splicing-dependent frameshifting
Precise and reliable gene delivery remains technically challenging. Here, the authors show that rationally designed frameshifting splicing can be used to express genes only in targeted cell types, with the potential to enhance the specificity AAV gene delivery.
- Jonathan P. Ling
- , Alexei M. Bygrave
- & Seth Blackshaw
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Article
| Open Access
Liver-directed lentiviral gene therapy corrects hemophilia A mice and achieves normal-range factor VIII activity in non-human primates
“Lentiviral gene therapy to the liver establishes stable long-term normal to supra-normal coagulation factor VIII activity in mouse models of hemophilia A and in non-human primates, representing a potential new treatment option for people with hemophilia A.”.
- Michela Milani
- , Cesare Canepari
- & Alessio Cantore
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Article
| Open Access
Subthalamic nucleus stabilizes movements by reducing neural spike variability in monkey basal ganglia
Chemogenetic inactivation of the subthalamic nucleus in monkeys increases spike train variability in the pallidum and prolongs movement time, suggesting its role in stabilizing pallidal spike trains to achieve stable motor control.
- Taku Hasegawa
- , Satomi Chiken
- & Atsushi Nambu
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Article
| Open Access
Choice of vector and surgical approach enables efficient cochlear gene transfer in nonhuman primate
Gene therapy using Adeno-associated viral vectors (AAV) rescues hearing and balance deficits in mouse models of human disorders. Here, the authors show that AAVAnc80L65 allows efficient cochlear gene transfer in nonhuman primates, and motivate future studies to evaluate gene therapy for hearing and balance disorders.
- Eva Andres-Mateos
- , Lukas D. Landegger
- & Luk H. Vandenberghe
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Article
| Open Access
A synthetic circuit for buffering gene dosage variation between individual mammalian cells
Transient plasmid transfection produces a wide distribution of copy numbers and expression heterogeneity. Here the authors present synthetic circuits called Equalizers that buffer copy number variation at the single-cell level and minimize cell-to-cell expression variability.
- Jin Yang
- , Jihwan Lee
- & François St-Pierre
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Article
| Open Access
Chemogenetic ON and OFF switches for RNA virus replication
Therapeutic application of RNA viruses requires tight control over viral activity. Here the authors design a regulatory switch that enables control over activity with clinically approved HIV protease inhibitors.
- E. Heilmann
- , J. Kimpel
- & D. von Laer
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Article
| Open Access
Engineering monocyte/macrophage−specific glucocerebrosidase expression in human hematopoietic stem cells using genome editing
Gaucher disease is a lysosomal storage disorder caused by insufficient glucocerebrosidase expression. Here, the authors describe a CRISPR/Cas9-based gene-editing approach to re-express this enzyme in human blood stem cells and show that they can engraft in NSG mice and differentiate into functional macrophages.
- Samantha G. Scharenberg
- , Edina Poletto
- & Natalia Gomez-Ospina
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Article
| Open Access
Caliciviral protein-based artificial translational activator for mammalian gene circuits with RNA-only delivery
Synthetic RNA-based circuits allow for gene regulation without the risk of mutagenesis. Here the authors demonstrate a Caliciviral VPg-based Translational activator which allows translational activation of synthetic mRNAs.
- Hideyuki Nakanishi
- & Hirohide Saito
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Article
| Open Access
Gene therapy for progressive familial intrahepatic cholestasis type 3 in a clinically relevant mouse model
Progressive familial intrahepatic cholestasis type 3 is a disease caused by mutations in the gene encoding the multidrug resistance protein 3, and has limited treatment options. Here they show that adeno-associated virus mediated gene therapy prevents disease progression in a mouse disease model.
- Nicholas D. Weber
- , Leticia Odriozola
- & Cristian Smerdou
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| Open Access
AAV-ie enables safe and efficient gene transfer to inner ear cells
There are currently few AAV vectors that can effectively target the diverse cell types of the inner ear. Here the authors design AAV-ie for gene delivery to the mouse cochlea, targeting cochlear supporting cells.
- Fangzhi Tan
- , Cenfeng Chu
- & Guisheng Zhong
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Article
| Open Access
An anaerobic bacterium host system for heterologous expression of natural product biosynthetic gene clusters
Anaerobic bacteria represent a rich source of biological and chemical diversity but are difficult to cultivate and there is a lack of heterologous expression systems. Here the authors develop an expression system based on S. mutans UA159 for biosynthetic gene clusters from anaerobic bacteria.
- Tingting Hao
- , Zhoujie Xie
- & Yihua Chen
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Article
| Open Access
Sub-2 Å Ewald curvature corrected structure of an AAV2 capsid variant
Single-particle cryo-EM is a powerful method for macromolecular structure determination. Here the authors demonstrate that Ewald sphere curvature correction, sub-Angstrom pixilation and per-particle CTF refinement can improve map quality and resolution and present the 1.86 Å cryo-EM structure of an adeno-associated virus serotype 2 variant.
- Yong Zi Tan
- , Sriram Aiyer
- & Dmitry Lyumkis
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Article
| Open Access
Engineered bidirectional promoters enable rapid multi-gene co-expression optimization
Classic monodirectional promoters are of limited use for multiple gene co-expression. Here the authors generate a library of 168 bidirectional promoters for the yeast K. phaffii (syn. P. pastoris) with diverse expression profiles to optimize metabolic pathway design.
- Thomas Vogl
- , Thomas Kickenweiz
- & Anton Glieder
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Article
| Open Access
AAV vector-mediated in vivo reprogramming into pluripotency
In vivo reprogramming of somatic cells is hampered by the need for vectors to express the OKSM factors in selected organs. Here the authors report new AAV-based vectors capable of in vivo reprogramming at low doses.
- Elena Senís
- , Lluc Mosteiro
- & Dirk Grimm
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Article
| Open Access
Adenoviral vector with shield and adapter increases tumor specificity and escapes liver and immune control
Viral gene therapy can be limited by the efficacy of virion sequestration, immune responses and the silencing of genetic payloads. Here the authors engineer an advenovirus protein coat which shields the virion from the immune system while targeting cancer cells.
- Markus Schmid
- , Patrick Ernst
- & Andreas Plückthun
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Article
| Open Access
PUMA gene delivery to synoviocytes reduces inflammation and degeneration of arthritic joints
Proliferation of synoviocytes contributes to joint damage in rheumatoid arthritis. Here the authors show that targeting of these cells by a vector, consisting of a baculovirus conjugated to an adenovirus carrying the pro-apoptotic gene PUMA, has therapeutic efficacy in a rat arthritis model.
- Saw-See Hong
- , Hubert Marotte
- & Pierre Miossec
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Article
| Open Access
A robot for high yield electrophysiology and morphology of single neurons in vivo
Single-cell characterization and perturbation of neurons is critical for revealing the structure-function relationship of brain cells. Here the authors develop a robot that performs single-cell electroporation and extracellular electrophysiology and can be used for performingin vivosingle-cell experiments in deep brain tissues optically difficult to access.
- Lu Li
- , Benjamin Ouellette
- & Hongkui Zeng
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Article
| Open Access
Enhancing titres of therapeutic viral vectors using the transgene repression in vector production (TRiP) system
The maximum titre of therapeutic viral vectors can be adversely affected by the encoded transgene. Here the authors repress transgene expression in producing cells by employing the tryptophan RNA-binding attenuation protein and show that it improves titre of RNA- and DNA-based viral vectors expressing toxic transgenes.
- H. E. Maunder
- , J. Wright
- & D. C. Farley
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| Open Access
Highly efficient baculovirus-mediated multigene delivery in primary cells
Current viral gene delivery systems are limited in the amount of foreign DNA they can deliver to cells. Here the authors develop MultiPrime, a baculovirus-based vector system capable of multigene delivery into a wide variety of cells, and use Multiprime for genome engineering by CRISPR/Cas9.
- Maysam Mansouri
- , Itxaso Bellon-Echeverria
- & Philipp Berger
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Article
| Open Access
Designer diatom episomes delivered by bacterial conjugation
Algae hold great promise for biofuel and chemical production but their use as model systems is hampered by the absence of suitable genetic tools. Here Karas et al. present a nuclear episomal vector for diatoms that is maintained in the absence of antibiotics, and a plasmid delivery method via conjugation with E. coli.
- Bogumil J. Karas
- , Rachel E. Diner
- & Philip D. Weyman
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Off-target-free gene delivery by affinity-purified receptor-targeted viral vectors
The clinical use of adeno-associated virus vectors (AAVs) has been limited by the lack of transduction specificity. Here the authors show that receptor-targeted, affinity-tagged, and purified AVVs reach tumours in mouse models with high selectivity and efficiency, outperforming therapeutic antibodies.
- Robert C. Münch
- , Anke Muth
- & Christian J. Buchholz
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A fluorinated dendrimer achieves excellent gene transfection efficacy at extremely low nitrogen to phosphorus ratios
Polymers represent promising gene vectors due to their high efficiency and low cytotoxicity. Here, the authors show that fluorination increases gene transfection efficacy, while reducing cytotoxicity, and suggest an important role for this strategy in the design of efficient gene vectors.
- Mingming Wang
- , Hongmei Liu
- & Yiyun Cheng
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A plasmid-based multigene expression system for mammalian cells
Fluorescent proteins are widely used in molecular biology to visualize protein expression and localization. Here, Krizet al. describe an efficient and flexible modular plasmid-based eukaryotic expression cloning strategy for the homogeneous expression of several fluorescent proteins in one cell.
- Andrijana Kriz
- , Katharina Schmid
- & Philipp Berger
A toolbox of astrocyte-specific, serotype-independent adeno-associated viral vectors using microRNA targeting sequences