Messenger RNA Delivery for Tissue Engineering and Regenerative Medicine Applications.

The ability to control cellular processes and precisely direct cellular reprogramming has revolutionized regenerative medicine. Recent advances in in vitro transcribed (IVT) mRNA technology with chemical modifications have led to development of methods that control spatiotemporal gene expression. Additionally, there is a current thrust toward the development of safe, integration-free approaches to gene therapy for translational purposes. In this review, we describe strategies of synthetic IVT mRNA modifications and nonviral technologies for intracellular delivery. We provide insights into the current tissue engineering approaches that use a hydrogel scaffold with genetic material. Furthermore, we discuss the transformative potential of novel mRNA formulations that when embedded in hydrogels can trigger controlled genetic manipulation to regenerate tissues and organs in vitro and in vivo. The role of mRNA delivery in vascularization, cytoprotection, and Cas9-mediated xenotransplantation is additionally highlighted. Harmonizing mRNA delivery vehicle interactions with polymeric scaffolds can be used to present genetic cues that lead to precise command over cellular reprogramming, differentiation, and secretome activity of stem cells-an ultimate goal for tissue engineering.

[1]  Robert Langer,et al.  Lipid Nanoparticle Assisted mRNA Delivery for Potent Cancer Immunotherapy. , 2017, Nano letters.

[2]  Daniel G. Anderson,et al.  Therapeutic genome editing by combined viral and non-viral delivery of CRISPR system components in vivo , 2016, Nature Biotechnology.

[3]  Jimin Gao,et al.  Efficient generation of gene-modified pigs via injection of zygote with Cas9/sgRNA , 2015, Scientific Reports.

[4]  Wim E Hennink,et al.  25th Anniversary Article: Engineering Hydrogels for Biofabrication , 2013, Advanced materials.

[5]  Daniel Anderson,et al.  Delivery materials for siRNA therapeutics. , 2013, Nature materials.

[6]  P. Edwards,et al.  Sonic hedgehog gene-enhanced tissue engineering for bone regeneration , 2005, Gene Therapy.

[7]  Robert Lanza,et al.  Generation of human induced pluripotent stem cells by direct delivery of reprogramming proteins. , 2009, Cell stem cell.

[8]  Hiroshi Yagi,et al.  Organ reengineering through development of a transplantable recellularized liver graft using decellularized liver matrix , 2010, Nature Medicine.

[9]  J. Wong,et al.  Reprogramming of somatic cells via TAT-mediated protein transduction of recombinant factors. , 2012, Biomaterials.

[10]  X. Qin,et al.  Paracrine action enhances the effects of autologous mesenchymal stem cell transplantation on vascular regeneration in rat model of myocardial infarction. , 2005, The Annals of thoracic surgery.

[11]  Jiannis Ragoussis,et al.  Direct reprogramming of fibroblasts into endothelial cells capable of angiogenesis and reendothelialization in tissue-engineered vessels , 2012, Proceedings of the National Academy of Sciences.

[12]  Mark Ibberson,et al.  Endogenous RNAs modulate microRNA sorting to exosomes and transfer to acceptor cells. , 2014, Cell reports.

[13]  P. Cullis,et al.  On the mechanism whereby cationic lipids promote intracellular delivery of polynucleic acids , 2001, Gene Therapy.

[14]  Jason A. Burdick,et al.  Engineered Hydrogels for Local and Sustained Delivery of RNA‐Interference Therapies , 2017, Advanced healthcare materials.

[15]  Jean-Louis Viovy,et al.  A review of microfabrication and hydrogel engineering for micro-organs on chips. , 2014, Biomaterials.

[16]  Kelly Servick,et al.  This mysterious $2 billion biotech is revealing the secrets behind its new drugs and vaccines , 2017 .

[17]  J. R. Vargas,et al.  Charge-altering releasable transporters (CARTs) for the delivery and release of mRNA in living animals , 2017, Proceedings of the National Academy of Sciences.

[18]  Sruthi Mantri,et al.  CRISPR/Cas9 β-globin gene targeting in human haematopoietic stem cells , 2016, Nature.

[19]  Jeffry D. Sander,et al.  CRISPR-Cas systems for editing, regulating and targeting genomes , 2014, Nature Biotechnology.

[20]  Daniel G Anderson,et al.  Polymer-Lipid Nanoparticles for Systemic Delivery of mRNA to the Lungs. , 2016, Angewandte Chemie.

[21]  P. Mali,et al.  Efficient human iPS cell derivation by a non-integrating plasmid from blood cells with unique epigenetic and gene expression signatures , 2011, Cell Research.

[22]  Masaki Ieda,et al.  Direct reprogramming of fibroblasts into functional cardiomyocytes by defined factors. , 2010, Cell.

[23]  Christopher B. Rives,et al.  Plasmid delivery in vivo from porous tissue-engineering scaffolds: transgene expression and cellular transfection. , 2005, Molecular therapy : the journal of the American Society of Gene Therapy.

[24]  Alexei A. Sharov,et al.  Database for mRNA Half-Life of 19 977 Genes Obtained by DNA Microarray Analysis of Pluripotent and Differentiating Mouse Embryonic Stem Cells , 2008, DNA research : an international journal for rapid publication of reports on genes and genomes.

[25]  V. Rotello,et al.  Photocleavable Hydrogels for Light‐Triggered siRNA Release , 2016, Advanced healthcare materials.

[26]  H. Rammensee,et al.  Intradermal vaccinations with RNA coding for TAA generate CD8+ and CD4+ immune responses and induce clinical benefit in vaccinated patients. , 2011, Molecular therapy : the journal of the American Society of Gene Therapy.

[27]  I. Chen,et al.  Induced pluripotent stem cell reprogramming by integration-free Sendai virus vectors from peripheral blood of patients with craniometaphyseal dysplasia. , 2013, Cellular reprogramming.

[28]  Jeffrey Wilusz,et al.  The highways and byways of mRNA decay , 2007, Nature Reviews Molecular Cell Biology.

[29]  T. D. de Gruijl,et al.  Functional delivery of viral miRNAs via exosomes , 2010, Proceedings of the National Academy of Sciences.

[30]  D. Ingber,et al.  Reconstituting Organ-Level Lung Functions on a Chip , 2010, Science.

[31]  K. Zatloukal,et al.  Influenza virus hemagglutinin HA-2 N-terminal fusogenic peptides augment gene transfer by transferrin-polylysine-DNA complexes: toward a synthetic virus-like gene-transfer vehicle. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[32]  Emmanouil D Karagiannis,et al.  Rational design of a biomimetic cell penetrating peptide library. , 2013, ACS nano.

[33]  K. Kataoka,et al.  Systemic delivery of messenger RNA for the treatment of pancreatic cancer using polyplex nanomicelles with a cholesterol moiety. , 2016, Biomaterials.

[34]  Christian Schuetz,et al.  Regeneration and orthotopic transplantation of a bioartificial lung , 2010, Nature Medicine.

[35]  Gaurav Sahay,et al.  Endocytosis of nanomedicines. , 2010, Journal of controlled release : official journal of the Controlled Release Society.

[36]  M. Hasegawa,et al.  Efficient induction of transgene-free human pluripotent stem cells using a vector based on Sendai virus, an RNA virus that does not integrate into the host genome , 2009, Proceedings of the Japan Academy. Series B, Physical and biological sciences.

[37]  B. Karlan,et al.  Directed Differentiation of Human Induced Pluripotent Stem Cells into Fallopian Tube Epithelium , 2017, Scientific Reports.

[38]  A. Brivanlou,et al.  An efficient and reversible transposable system for gene delivery and lineage-specific differentiation in human embryonic stem cells. , 2009, Cell stem cell.

[39]  A. Nagy,et al.  Transgene-free production of pluripotent stem cells using piggyBac transposons. , 2011, Methods in molecular biology.

[40]  Wei Wang,et al.  piggyBac transposition reprograms fibroblasts to induced pluripotent stem cells , 2009, Nature.

[41]  T. Fitzgerald,et al.  Induction of KLF4 in basal keratinocytes blocks the proliferation–differentiation switch and initiates squamous epithelial dysplasia , 2005, Oncogene.

[42]  Wei Wang,et al.  Cationic nanocarriers induce cell necrosis through impairment of Na+/K+-ATPase and cause subsequent inflammatory response , 2015, Cell Research.

[43]  Gaurav Sahay,et al.  Boosting Intracellular Delivery of Lipid Nanoparticle-Encapsulated mRNA. , 2017, Nano letters.

[44]  David Bryder,et al.  Efficient ablation of genes in human hematopoietic stem and effector cells using CRISPR/Cas9. , 2014, Cell stem cell.

[45]  Zhen W. Zhuang,et al.  Tissue-Engineered Lungs for in Vivo Implantation , 2010, Science.

[46]  Tao Jiang,et al.  Exosomes derived from Rab27a‑overexpressing tumor cells elicit efficient induction of antitumor immunity. , 2013, Molecular medicine reports.

[47]  Luigi Biancone,et al.  Endothelial progenitor cell derived microvesicles activate an angiogenic program in endothelial cells by a horizontal transfer of mRNA. , 2007, Blood.

[48]  M. Rubinstein,et al.  Mobility of Nonsticky Nanoparticles in Polymer Liquids. , 2011, Macromolecules.

[49]  A. Lamond,et al.  Multidimensional proteomics for cell biology , 2015, Nature Reviews Molecular Cell Biology.

[50]  C. Pichon,et al.  Lipid-based mRNA vaccine delivery systems , 2015, Expert review of vaccines.

[51]  D. Russell,et al.  Adeno-associated virus finds its disease , 2015, Nature Genetics.

[52]  S. Armes,et al.  Biomimetic stimulus-responsive star diblock gelators. , 2005, Langmuir : the ACS journal of surfaces and colloids.

[53]  B. Bettencourt,et al.  Efficacy and safety of patisiran for familial amyloidotic polyneuropathy: a phase II multi-dose study , 2015, Orphanet Journal of Rare Diseases.

[54]  Daniel G. Miller,et al.  AAV Vector Integration Sites in Mouse Hepatocellular Carcinoma , 2007, Science.

[55]  Mark E. Davis,et al.  Cyclodextrin-modified polyethylenimine polymers for gene delivery. , 2004, Bioconjugate chemistry.

[56]  Yuhua Wang,et al.  Combination Immunotherapy of MUC1 mRNA Nano-vaccine and CTLA-4 Blockade Effectively Inhibits Growth of Triple Negative Breast Cancer. , 2018, Molecular therapy : the journal of the American Society of Gene Therapy.

[57]  D W Hutmacher,et al.  Three-Dimensional Bioprinting for Regenerative Dentistry and Craniofacial Tissue Engineering , 2015, Journal of dental research.

[58]  Brian J Cummings,et al.  Plasmid releasing multiple channel bridges for transgene expression after spinal cord injury. , 2009, Molecular therapy : the journal of the American Society of Gene Therapy.

[59]  M. Mahkam.  Novel pH-sensitive hydrogels for colon-specific drug delivery , 2010, Drug delivery.

[60]  Kaoru Tominaga,et al.  Activation of innate immune antiviral response by NOD2 , 2009, Nature Immunology.

[61]  Wei Li,et al.  One-step generation of knockout pigs by zygote injection of CRISPR/Cas system , 2014, Cell Research.

[62]  U. Şahin,et al.  Modification of antigen-encoding RNA increases stability, translational efficacy, and T-cell stimulatory capacity of dendritic cells. , 2006, Blood.

[63]  Chaenyung Cha,et al.  25th Anniversary Article: Rational Design and Applications of Hydrogels in Regenerative Medicine , 2014, Advanced materials.

[64]  Sue Fletcher,et al.  Regulation of eukaryotic gene expression by the untranslated gene regions and other non-coding elements , 2012, Cellular and Molecular Life Sciences.

[65]  R Langer,et al.  Functional arteries grown in vitro. , 1999, Science.

[66]  Martin Fussenegger,et al.  Therapeutic protein transduction of mammalian cells and mice by nucleic acid-free lentiviral nanoparticles , 2006, Nucleic acids research.

[67]  M. Tuszynski,et al.  BDNF-expressing marrow stromal cells support extensive axonal growth at sites of spinal cord injury , 2005, Experimental Neurology.

[68]  T. Kissel,et al.  Delivery of messenger RNA using poly(ethylene imine)-poly(ethylene glycol)-copolymer blends for polyplex formation: biophysical characterization and in vitro transfection properties. , 2010, Journal of controlled release : official journal of the Controlled Release Society.

[69]  Daniel G. Anderson,et al.  Materials for non-viral intracellular delivery of messenger RNA therapeutics. , 2016, Journal of controlled release : official journal of the Controlled Release Society.

[70]  Tao Pan,et al.  RNA modifications and structures cooperate to guide RNA–protein interactions , 2017, Nature Reviews Molecular Cell Biology.

[71]  M. Penn,et al.  A Novel Role for CAMKK1 in the Regulation of the Mesenchymal Stem Cell Secretome , 2017, Stem cells translational medicine.

[72]  S. Fukushima,et al.  Messenger RNA delivery of a cartilage-anabolic transcription factor as a disease-modifying strategy for osteoarthritis treatment , 2016, Scientific Reports.

[73]  C. Fathman,et al.  A short pulse of IL-4 delivered by DCs electroporated with modified mRNA can both prevent and treat autoimmune diabetes in NOD mice. , 2010, Molecular therapy : the journal of the American Society of Gene Therapy.

[74]  Kevin Eggan,et al.  Conversion of mouse and human fibroblasts into functional spinal motor neurons. , 2011, Cell stem cell.

[75]  M. Ohtsuka,et al.  The combinational use of CRISPR/Cas9‐based gene editing and targeted toxin technology enables efficient biallelic knockout of the α‐1,3‐galactosyltransferase gene in porcine embryonic fibroblasts , 2014, Xenotransplantation.

[76]  J. Kowalska,et al.  Azido-Functionalized 5' Cap Analogues for the Preparation of Translationally Active mRNAs Suitable for Fluorescent Labeling in Living Cells. , 2017, Angewandte Chemie.

[77]  R. Cancedda,et al.  The Regenerative Role of the Fetal and Adult Stem Cell Secretome , 2013, Journal of clinical medicine.

[78]  K. G. Rajeev,et al.  Rational design of cationic lipids for siRNA delivery , 2010, Nature Biotechnology.

[79]  Dan Peer,et al.  Toxicity profiling of several common RNAi-based nanomedicines: a comparative study , 2013, Drug Delivery and Translational Research.

[80]  I. Verma,et al.  Systemic delivery of factor IX messenger RNA for protein replacement therapy , 2017, Proceedings of the National Academy of Sciences.

[81]  D. Wishart.  Emerging applications of metabolomics in drug discovery and precision medicine , 2016, Nature Reviews Drug Discovery.

[82]  Xin-Yuan Fu,et al.  STAT3 mediates bone marrow mesenchymal stem cell VEGF production. , 2007, Journal of molecular and cellular cardiology.

[83]  Shubiao Zhang,et al.  Toxicity of cationic lipids and cationic polymers in gene delivery. , 2006, Journal of controlled release : official journal of the Controlled Release Society.

[84]  D. Weissman,et al.  Zika virus protection by a single low dose nucleoside modified mRNA vaccination , 2017, Nature.

[85]  M. Park,et al.  Injectable polyplex hydrogel for localized and long-term delivery of siRNA. , 2012, ACS nano.

[86]  V. Hombach,et al.  mRNA-Mediated Gene Delivery Into Human Progenitor Cells Promotes Highly Efficient Protein Expression , 2007, Journal of cellular and molecular medicine.

[87]  M. Gale,et al.  Immune signaling by RIG-I-like receptors. , 2011, Immunity.

[88]  D. Weissman,et al.  mRNA: Fulfilling the Promise of Gene Therapy. , 2015, Molecular therapy : the journal of the American Society of Gene Therapy.

[89]  Kam W. Leong,et al.  Whole Blood Cells Loaded with Messenger RNA as an Anti‐Tumor Vaccine , 2014, Advanced healthcare materials.

[90]  Marius Wernig,et al.  c-Myc is dispensable for direct reprogramming of mouse fibroblasts. , 2008, Cell stem cell.

[91]  Jason S. Meyer,et al.  Robust Differentiation of mRNA-Reprogrammed Human Induced Pluripotent Stem Cells Toward a Retinal Lineage , 2016, Stem cells translational medicine.

[92]  H. Ouyang,et al.  Efficient Generation of Myostatin Mutations in Pigs Using the CRISPR/Cas9 System , 2015, Scientific Reports.

[93]  K. Woltjen,et al.  Virus free induction of pluripotency and subsequent excision of reprogramming factors , 2009, Nature.

[94]  Lei Zhang,et al.  VEGF/SDF-1 promotes cardiac stem cell mobilization and myocardial repair in the infarcted heart. , 2011, Cardiovascular Research.

[95]  Takahiro Nomoto,et al.  Modulated protonation of side chain aminoethylene repeats in N-substituted polyaspartamides promotes mRNA transfection. , 2014, Journal of the American Chemical Society.

[96]  Kerstin F. Gerer,et al.  Electroporation of mRNA as Universal Technology Platform to Transfect a Variety of Primary Cells with Antigens and Functional Proteins. , 2017, Methods in molecular biology.

[97]  Hiroki Kato,et al.  Incorporation of pseudouridine into mRNA yields superior nonimmunogenic vector with increased translational capacity and biological stability. , 2008, Molecular therapy : the journal of the American Society of Gene Therapy.

[98]  D. Ingber,et al.  Microfluidic organs-on-chips , 2014, Nature Biotechnology.

[99]  Michel Sadelain,et al.  Stoichiometric and temporal requirements of Oct4, Sox2, Klf4, and c-Myc expression for efficient human iPSC induction and differentiation , 2009, Proceedings of the National Academy of Sciences.

[100]  T. Ichisaka,et al.  Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined Factors , 2007, Cell.

[101]  Jeffrey M Karp,et al.  Engineering Stem Cell Organoids. , 2016, Cell stem cell.

[102]  H. Dvorak,et al.  Vascular hyperpermeability, angiogenesis, and stroma generation. , 2012, Cold Spring Harbor perspectives in medicine.

[103]  C. Limatola,et al.  Differentiation of control and ALS mutant human iPSCs into functional skeletal muscle cells, a tool for the study of neuromuscolar diseases , 2016, Stem cell research.

[104]  Lei Wang,et al.  Generation of Gene-Modified Cynomolgus Monkey via Cas9/RNA-Mediated Gene Targeting in One-Cell Embryos , 2014, Cell.

[105]  Y. Barenholz.  Doxil®--the first FDA-approved nano-drug: lessons learned. , 2012, Journal of controlled release : official journal of the Controlled Release Society.

[106]  B. Feeley,et al.  Lentivirus-mediated gene transfer induces long-term transgene expression of BMP-2 in vitro and new bone formation in vivo. , 2005, Molecular therapy : the journal of the American Society of Gene Therapy.

[107]  S. J. Coleman,et al.  Determinants and implications of mRNA poly(A) tail size--does this protein make my tail look big? , 2014, Seminars in cell & developmental biology.

[108]  R. Deering,et al.  Nucleic acid vaccines: prospects for non-viral delivery of mRNA vaccines , 2014, Expert opinion on drug delivery.

[109]  P. Bianco,et al.  Stem cells in tissue engineering , 2001, Nature.

[110]  Özlem Türeci,et al.  mRNA-based therapeutics — developing a new class of drugs , 2014, Nature Reviews Drug Discovery.

[111]  E. Alsberg,et al.  Sustained localized presentation of RNA interfering molecules from in situ forming hydrogels to guide stem cell osteogenic differentiation. , 2014, Biomaterials.

[112]  Yunlong Liu,et al.  Platelet-derived growth factor-BB enhances MSC-mediated cardioprotection via suppression of miR-320 expression. , 2015, American journal of physiology. Heart and circulatory physiology.

[113]  D. Givol,et al.  Reprogramming of human fibroblasts to pluripotent stem cells using mRNA of four transcription factors. , 2010, Biochemical and biophysical research communications.

[114]  D. Mooney,et al.  Bone regeneration in a rat cranial defect with delivery of PEI-condensed plasmid DNA encoding for bone morphogenetic protein-4 (BMP-4) , 2005, Gene Therapy.

[115]  R. David,et al.  Defining Optimized Properties of Modified mRNA to Enhance Virus- and DNA- Independent Protein Expression in Adult Stem Cells and Fibroblasts , 2015, Cellular Physiology and Biochemistry.

[116]  Nick Barker,et al.  Organoids as an in vitro model of human development and disease , 2016, Nature Cell Biology.

[117]  M. Ogris,et al.  Peptide-mediated RNA delivery: a novel approach for enhanced transfection of primary and post-mitotic cells. , 2001, Nucleic acids research.

[118]  Houping Ni,et al.  Suppression of RNA recognition by Toll-like receptors: the impact of nucleoside modification and the evolutionary origin of RNA. , 2005, Immunity.

[119]  S. Akira,et al.  Species-Specific Recognition of Single-Stranded RNA via Toll-like Receptor 7 and 8 , 2004, Science.

[120]  Mehmet Fatih Yanik,et al.  mRNA-engineered mesenchymal stem cells for targeted delivery of interleukin-10 to sites of inflammation. , 2013, Blood.

[121]  P. Burridge,et al.  Generation of human iPSCs from human peripheral blood mononuclear cells using non-integrative Sendai virus in chemically defined conditions. , 2013, Methods in molecular biology.

[122]  M. Park,et al.  An injectable cell penetrable nano-polyplex hydrogel for localized siRNA delivery. , 2013, Biomaterials.

[123]  S. Yamanaka,et al.  Induction of Pluripotent Stem Cells from Mouse Embryonic and Adult Fibroblast Cultures by Defined Factors , 2006, Cell.

[124]  P. Linsley,et al.  Recognizing and avoiding siRNA off-target effects for target identification and therapeutic application , 2010, Nature Reviews Drug Discovery.

[125]  Shinya Yamanaka,et al.  Generation of Mouse Induced Pluripotent Stem Cells Without Viral Vectors , 2008, Science.

[126]  J. Utikal,et al.  Induced Pluripotent Stem Cells Generated Without Viral Integration , 2008, Science.

[127]  Hong Cao,et al.  Guanabenz (Wytensin™) selectively enhances uptake and efficacy of hydrophobically modified siRNAs , 2015, Nucleic acids research.

[128]  Jiankui Zhou,et al.  Dual sgRNAs facilitate CRISPR/Cas9‐mediated mouse genome targeting , 2014, The FEBS journal.

[129]  T. Kissel,et al.  Efficient and Tumor Targeted siRNA Delivery by Polyethylenimine-graft-polycaprolactone-block-poly(ethylene glycol)-folate (PEI-PCL-PEG-Fol). , 2016, Molecular pharmaceutics.

[130]  Meifeng Xu,et al.  Exosomes secreted from GATA-4 overexpressing mesenchymal stem cells serve as a reservoir of anti-apoptotic microRNAs for cardioprotection. , 2015, International journal of cardiology.

[131]  Justin M. Richner,et al.  Modified mRNA Vaccines Protect against Zika Virus Infection , 2017, Cell.

[132]  N. Sonenberg,et al.  N1-methyl-pseudouridine in mRNA enhances translation through eIF2α-dependent and independent mechanisms by increasing ribosome density , 2017, Nucleic acids research.

[133]  Zhisong He,et al.  Paracrine factors released by GATA-4 overexpressed mesenchymal stem cells increase angiogenesis and cell survival. , 2010, American journal of physiology. Heart and circulatory physiology.

[134]  M. Tector,et al.  Evaluation of human and non‐human primate antibody binding to pig cells lacking GGTA1/CMAH/β4GalNT2 genes , 2015, Xenotransplantation.

[135]  Rui-Zhen Shi,et al.  Angiotensin II induces vascular endothelial growth factor synthesis in mesenchymal stem cells. , 2009, Experimental cell research.

[136]  Eben Alsberg,et al.  Light-triggered RNA release and induction of hMSC osteogenesis via photodegradable, dual-crosslinked hydrogels. , 2016, Nanomedicine.

[137]  Markus J Buehler,et al.  Materiomics: An ‐omics Approach to Biomaterials Research , 2013, Advanced materials.

[138]  Anandika Dhaliwal,et al.  Transfection in the third dimension. , 2013, Integrative biology : quantitative biosciences from nano to macro.

[139]  K. Kataoka,et al.  Messenger RNA-based therapeutics for the treatment of apoptosis-associated diseases , 2015, Scientific Reports.

[140]  S. Lim,et al.  Exosomes for drug delivery - a novel application for the mesenchymal stem cell. , 2013, Biotechnology advances.

[141]  A. Bosio,et al.  Direct and efficient transfection of mouse neural stem cells and mature neurons by in vivo mRNA electroporation , 2017, Development.

[142]  Haihua Xiao,et al.  A Targeted and Stable Polymeric Nanoformulation Enhances Systemic Delivery of mRNA to Tumors. , 2017, Molecular therapy : the journal of the American Society of Gene Therapy.

[143]  A. Schnerch,et al.  Direct conversion of human fibroblasts to multilineage blood progenitors , 2010, Nature.

[144]  Tae-Jin Lee,et al.  Efficient mRNA delivery with graphene oxide-polyethylenimine for generation of footprint-free human induced pluripotent stem cells. , 2016, Journal of controlled release : official journal of the Controlled Release Society.

[145]  J. Hecker,et al.  Lipid-mediated delivery of RNA is more efficient than delivery of DNA in non-dividing cells. , 2010, International journal of pharmaceutics.

[146]  R. Rhoads,et al.  Novel "anti-reverse" cap analogs with superior translational properties. , 2003, RNA.

[147]  Renhui Yang,et al.  Exaggerated Hypotensive Effect of Vascular Endothelial Growth Factor in Spontaneously Hypertensive Rats , 2002, Hypertension.

[148]  David W. McComb,et al.  An Orthogonal Array Optimization of Lipid-like Nanoparticles for mRNA Delivery in Vivo. , 2015, Nano letters.

[149]  Rudolf Jaenisch,et al.  Ectopic Expression of Oct-4 Blocks Progenitor-Cell Differentiation and Causes Dysplasia in Epithelial Tissues , 2005, Cell.

[150]  B. Rabie,et al.  Electroporation for Transfection and Differentiation of Dental Pulp Stem Cells , 2013, BioResearch open access.

[151]  Steffen Loft,et al.  In vivo toxicity of cationic micelles and liposomes. , 2015, Nanomedicine : nanotechnology, biology, and medicine.

[152]  Junmin Zhu,et al.  Bioactive modification of poly(ethylene glycol) hydrogels for tissue engineering. , 2010, Biomaterials.

[153]  K. Jensen,et al.  In vitro and ex vivo strategies for intracellular delivery , 2016, Nature.

[154]  J. Holland,et al.  Elevated serum ribonuclease in patients with pancreatic cancer. , 1976, Proceedings of the National Academy of Sciences of the United States of America.

[155]  Takashi Aoi,et al.  Generation of induced pluripotent stem cells without Myc from mouse and human fibroblasts , 2008, Nature Biotechnology.

[156]  Y. Kan,et al.  Blood Cell‐Derived Induced Pluripotent Stem Cells Free of Reprogramming Factors Generated by Sendai Viral Vectors , 2013, Stem cells translational medicine.

[157]  D. Weissman,et al.  Increased Erythropoiesis in Mice Injected With Submicrogram Quantities of Pseudouridine-containing mRNA Encoding Erythropoietin , 2012, Molecular therapy : the journal of the American Society of Gene Therapy.

[158]  J. Burdick,et al.  Materials science: Radicals promote magnetic gel assembly , 2014, Nature.

[159]  N. Zhang,et al.  One-step generation of different immunodeficient mice with multiple gene modifications by CRISPR/Cas9 mediated genome engineering. , 2014, The international journal of biochemistry & cell biology.

[160]  B. Stoddard,et al.  Hit-and-run programming of therapeutic cytoreagents using mRNA nanocarriers , 2017, Nature Communications.

[161]  Soo-Chang Song,et al.  Targetable micelleplex hydrogel for long-term, effective, and systemic siRNA delivery. , 2014, Biomaterials.

[162]  Gunther Hartmann,et al.  5'-Triphosphate RNA Is the Ligand for RIG-I , 2006, Science.

[163]  Gaurav Sahay,et al.  Challenges in carrier-mediated intracellular delivery: moving beyond endosomal barriers. , 2016, Wiley interdisciplinary reviews. Nanomedicine and nanobiotechnology.

[164]  E. Lander,et al.  Development and Applications of CRISPR-Cas9 for Genome Engineering , 2014, Cell.

[165]  K. Mamchaoui,et al.  Synthetically modified mRNA for efficient and fast human iPS cell generation and direct transdifferentiation to myoblasts. , 2016, Biochemical and biophysical research communications.

[166]  K. Kataoka,et al.  Screening of mRNA Chemical Modification to Maximize Protein Expression with Reduced Immunogenicity , 2015, Pharmaceutics.

[167]  Anthony Atala,et al.  3D bioprinting of tissues and organs , 2014, Nature Biotechnology.

[168]  E. Alsberg,et al.  Functionalized, biodegradable hydrogels for control over sustained and localized siRNA delivery to incorporated and surrounding cells. , 2013, Acta biomaterialia.

[169]  Robert Langer,et al.  Genetic engineering of human stem cells for enhanced angiogenesis using biodegradable polymeric nanoparticles , 2009, Proceedings of the National Academy of Sciences.

[170]  S. Dowdy.  Overcoming cellular barriers for RNA therapeutics , 2017, Nature Biotechnology.

[171]  David A. Scott,et al.  Genome-wide binding of the CRISPR endonuclease Cas9 in mammalian cells , 2014, Nature Biotechnology.

[172]  Rudolf Jaenisch,et al.  One-Step Generation of Mice Carrying Mutations in Multiple Genes by CRISPR/Cas-Mediated Genome Engineering , 2013, Cell.

[173]  Wade Wang,et al.  Structurally Programmed Assembly of Translation Initiation Nanoplex for Superior mRNA Delivery. , 2017, ACS nano.

[174]  J. Ingwall,et al.  Evidence supporting paracrine hypothesis for Akt‐modified mesenchymal stem cell‐mediated cardiac protection and functional improvement , 2006, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[175]  Dong Wook Han,et al.  Generation of induced pluripotent stem cells using recombinant proteins. , 2009, Cell stem cell.

[176]  Bin Li,et al.  Effects of Chemically Modified Messenger RNA on Protein Expression. , 2016, Bioconjugate chemistry.

[177]  Xi-rong Guo,et al.  Feeder-Free Derivation of Human Induced Pluripotent Stem Cells with Messenger RNA , 2012, Scientific Reports.

[178]  David Y B Deng,et al.  Hypoxia-inducible factor 1-α-AA-modified bone marrow stem cells protect PC12 cells from hypoxia-induced apoptosis, partially through VEGF/PI3K/Akt/FoxO1 pathway. , 2012, Stem cells and development.

[179]  J. Kjems,et al.  Chitosan Hydrogel as siRNA vector for prolonged gene silencing , 2014, Journal of Nanobiotechnology.

[180]  Francesco Stellacci,et al.  Effect of surface properties on nanoparticle-cell interactions. , 2010, Small.

[181]  Khalid A. Hajj,et al.  Tools for translation: non-viral materials for therapeutic mRNA delivery , 2017 .

[182]  G. J. Dimitriadis.  Translation of rabbit globin mRNA introduced by liposomes into mouse lymphocytes , 1978, Nature.

[183]  A. Khademhosseini,et al.  Synthesis, properties, and biomedical applications of gelatin methacryloyl (GelMA) hydrogels. , 2015, Biomaterials.

[184]  Alexander Meissner,et al.  Highly efficient reprogramming to pluripotency and directed differentiation of human cells with synthetic modified mRNA. , 2010, Cell stem cell.

[185]  J. Walder,et al.  Substrate specificity and kinetics of degradation of antisense oligonucleotides by a 3' exonuclease in plasma. , 1991, Antisense research and development.

[186]  Roman Kaliszan,et al.  Metabolomics for laboratory diagnostics. , 2015, Journal of pharmaceutical and biomedical analysis.

[187]  J. Rosenecker,et al.  Expression of therapeutic proteins after delivery of chemically modified mRNA in mice , 2011, Nature Biotechnology.

[188]  Xin Li,et al.  A comparison of non-integrating reprogramming methods , 2014, Nature Biotechnology.

[189]  Ronald A. Li,et al.  Modified mRNA directs the fate of heart progenitor cells and induces vascular regeneration after myocardial infarction , 2013, Nature Biotechnology.

[190]  B. Spiegelman,et al.  ADD1/SREBP1 promotes adipocyte differentiation and gene expression linked to fatty acid metabolism. , 1996, Genes & development.

[191]  K. Miller,et al.  Self-adjuvanted mRNA vaccination in advanced prostate cancer patients: a first-in-man phase I/IIa study , 2015, Journal of Immunotherapy for Cancer.

[192]  Chuan He,et al.  Post-transcriptional gene regulation by mRNA modifications , 2016, Nature Reviews Molecular Cell Biology.

[193]  Hao Zhu,et al.  Non-Viral CRISPR/Cas Gene Editing In Vitro and In Vivo Enabled by Synthetic Nanoparticle Co-Delivery of Cas9 mRNA and sgRNA. , 2017, Angewandte Chemie.

[194]  H. Niemann,et al.  Efficient production of biallelic GGTA1 knockout pigs by cytoplasmic microinjection of CRISPR/Cas9 into zygotes , 2016, Xenotransplantation.

[195]  J. A. Bernal.  RNA-Based Tools for Nuclear Reprogramming and Lineage-Conversion: Towards Clinical Applications , 2013, Journal of Cardiovascular Translational Research.

[196]  Shizuo Akira,et al.  Innate Antiviral Responses by Means of TLR7-Mediated Recognition of Single-Stranded RNA , 2004, Science.

[197]  H. Niemann,et al.  The production of multi-transgenic pigs: update and perspectives for xenotransplantation , 2016, Transgenic Research.

[198]  T. Schoeb,et al.  Polycistronic Lentiviral Vector for “Hit and Run” Reprogramming of Adult Skin Fibroblasts to Induced Pluripotent Stem Cells , 2009, Stem cells.

[199]  D. Scherman,et al.  A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[200]  Mengrou Lu,et al.  Mesenchymal stem cells engineered to express selectin ligands and IL-10 exert enhanced therapeutic efficacy in murine experimental autoimmune encephalomyelitis. , 2016, Biomaterials.

[201]  M. Tector,et al.  Efficient generation of genetically distinct pigs in a single pregnancy using multiplexed single‐guide RNA and carbohydrate selection , 2015, Xenotransplantation.

[202]  G. Church,et al.  Supplementary Materials for Genome-wide inactivation of porcine endogenous retroviruses ( PERVs ) , 2015 .

[203]  Jianhui Gong,et al.  Correction of a pathogenic gene mutation in human embryos , 2017, Nature.

[204]  Jason P. Gleghorn,et al.  Microfluidic scaffolds for tissue engineering. , 2007, Nature materials.

[205]  Kimberly J. Hassett,et al.  Preclinical and Clinical Demonstration of Immunogenicity by mRNA Vaccines against H10N8 and H7N9 Influenza Viruses , 2017, Molecular therapy : the journal of the American Society of Gene Therapy.

[206]  F. W. Buaas,et al.  Delivery of Cas9 Protein into Mouse Zygotes through a Series of Electroporation Dramatically Increases the Efficiency of Model Creation. , 2016, Journal of genetics and genomics = Yi chuan xue bao.

[207]  D. B. Carter,et al.  Phenotyping of transgenic cloned piglets. , 2002, Cloning and stem cells.

[208]  H. Lee,et al.  Emergence of synthetic mRNA: In vitro synthesis of mRNA and its applications in regenerative medicine. , 2018, Biomaterials.

[209]  G. B. Robb,et al.  mRNA capping: biological functions and applications , 2016, Nucleic acids research.

[210]  D. Tollervey,et al.  The Many Pathways of RNA Degradation , 2009, Cell.

[211]  Ronald A. Li,et al.  Driving vascular endothelial cell fate of human multipotent Isl1+ heart progenitors with VEGF modified mRNA , 2013, Cell Research.

[212]  Pei-Rong Wang,et al.  Induction of hepatocellular carcinoma by in vivo gene targeting , 2012, Proceedings of the National Academy of Sciences.

[213]  Jeffrey Bonadio,et al.  Localized, direct plasmid gene delivery in vivo: prolonged therapy results in reproducible tissue regeneration , 1999, Nature Medicine.