Advances in CRISPR/Cas9 Technology for in Vivo Translation.

Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) technology has revolutionized therapeutic gene editing by providing researchers with a new method to study and cure diseases previously considered untreatable. While the full range and power of CRISPR technology for therapeutics is being elucidated through in vitro studies, translation to in vivo studies is slow. To date there is no totally effective delivery strategy to carry CRISPR components to the target site in vivo. The complexity of in vivo delivery is furthered by the number of potential delivery methods, the different forms in which CRISPR can be delivered as a therapeutic, and the disease target and tissue type in question. There are major challenges and limitations to delivery strategies, and it is imperative that future directions are guided by well-conducted studies that consider the full effect these variables have on the eventual outcome. In this review we will discuss the advances of the latest in vivo CRISPR/Cas9 delivery strategies and highlight the challenges yet to be overcome.

[1]  J. Wilson,et al.  Gene therapy: adenovirus vectors. , 1993, Current opinion in genetics & development.

[2]  Freeman Lan,et al.  Nanoparticle delivery of Cas9 ribonucleoprotein and donor DNA in vivo induces homology-directed DNA repair , 2017, Nature Biomedical Engineering.

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

[4]  D. Harki,et al.  SN-38 Conjugated Gold Nanoparticles Activated by Ewing Sarcoma Specific mRNAs Exhibit In Vitro and In Vivo Efficacy. , 2018, Bioconjugate chemistry.

[5]  Gang Bao,et al.  A high-fidelity Cas9 mutant delivered as a ribonucleoprotein complex enables efficient gene editing in human haematopoietic stem and progenitor cells , 2018, Nature Medicine.

[6]  Vincent M Rotello,et al.  Functionalized gold nanoparticles for drug delivery. , 2007, Nanomedicine.

[7]  Daniel G. Anderson,et al.  Non-viral vectors for gene-based therapy , 2014, Nature Reviews Genetics.

[8]  David R. Liu,et al.  CRISPR-Based Technologies for the Manipulation of Eukaryotic Genomes , 2017, Cell.

[9]  J. Doudna,et al.  The new frontier of genome engineering with CRISPR-Cas9 , 2014, Science.

[10]  B. Kinsey,et al.  Gene transfer by guanidinium‐cholesterol: dioleoylphosphatidyl‐ethanolamine liposome‐DNA complexes in aerosol , 1999, The journal of gene medicine.

[11]  T. Cheng,et al.  Conditional internalization of PEGylated nanomedicines by PEG engagers for triple negative breast cancer therapy , 2017, Nature Communications.

[12]  Mazhar Adli,et al.  The CRISPR tool kit for genome editing and beyond , 2018, Nature Communications.

[13]  John M. Shelton,et al.  Postnatal genome editing partially restores dystrophin expression in a mouse model of muscular dystrophy , 2016, Science.

[14]  C. Barbas,et al.  ZFN, TALEN, and CRISPR/Cas-based methods for genome engineering. , 2013, Trends in biotechnology.

[15]  D. Scherman,et al.  Plasmid DNA size does not affect the physicochemical properties of lipoplexes but modulates gene transfer efficiency. , 1999, Nucleic acids research.

[16]  J. Wolff,et al.  Breaking the bonds: non-viral vectors become chemically dynamic. , 2008, Molecular therapy : the journal of the American Society of Gene Therapy.

[17]  S. Takeoka,et al.  Plasmid DNA-encapsulating liposomes: effect of a spacer between the cationic head group and hydrophobic moieties of the lipids on gene expression efficiency. , 2009, Biochimica et biophysica acta.

[18]  Lukas E Dow,et al.  Inducible in vivo genome editing with CRISPR/Cas9 , 2015, Nature Biotechnology.

[19]  G. Church,et al.  Cas9 as a versatile tool for engineering biology , 2013, Nature Methods.

[20]  Hao Yin,et al.  Genome editing with Cas9 in adult mice corrects a disease mutation and phenotype , 2014, Nature Biotechnology.

[21]  Yanhua Rao,et al.  Strategies for controlling CRISPR/Cas9 off-target effects and biological variations in mammalian genome editing experiments. , 2018, Journal of biotechnology.

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

[23]  Zhen Gu,et al.  Macrophage-Specific in Vivo Gene Editing Using Cationic Lipid-Assisted Polymeric Nanoparticles. , 2018, ACS nano.

[24]  Lu Zhang,et al.  Efficient precise knockin with a double cut HDR donor after CRISPR/Cas9-mediated double-stranded DNA cleavage , 2017, Genome Biology.

[25]  Feng Zhang,et al.  In vivo interrogation of gene function in the mammalian brain using CRISPR-Cas9 , 2014, Nature Biotechnology.

[26]  Joshua J. Breunig,et al.  In Vivo CRISPR/Cas9 Gene Editing Corrects Retinal Dystrophy in the S334ter-3 Rat Model of Autosomal Dominant Retinitis Pigmentosa , 2015, Molecular therapy : the journal of the American Society of Gene Therapy.

[27]  Chao Yang,et al.  Adenovirus-mediated gene delivery: Potential applications for gene and cell-based therapies in the new era of personalized medicine , 2017, Genes & diseases.

[28]  Yang Yang,et al.  A dual AAV system enables the Cas9-mediated correction of a metabolic liver disease in newborn mice , 2016, Nature Biotechnology.

[29]  Hao Yin,et al.  Delivery technologies for genome editing , 2017, Nature Reviews Drug Discovery.

[30]  T. Reineke,et al.  Membrane and nuclear permeabilization by polymeric pDNA vehicles: efficient method for gene delivery or mechanism of cytotoxicity? , 2012, Molecular Pharmaceutics.

[31]  F. Cocchiarella,et al.  In vivo Editing of the Human Mutant Rhodopsin Gene by Electroporation of Plasmid-based CRISPR/Cas9 in the Mouse Retina , 2016, Molecular therapy. Nucleic acids.

[32]  Theresa A. Storm,et al.  The host response to adenovirus, helper-dependent adenovirus, and adeno-associated virus in mouse liver. , 2008, Molecular therapy : the journal of the American Society of Gene Therapy.

[33]  David A. Scott,et al.  Double Nicking by RNA-Guided CRISPR Cas9 for Enhanced Genome Editing Specificity , 2013, Cell.

[34]  Daniel G. Anderson,et al.  Knocking down barriers: advances in siRNA delivery , 2009, Nature Reviews Drug Discovery.

[35]  J. Doudna,et al.  Expanding the Biologist's Toolkit with CRISPR-Cas9. , 2015, Molecular cell.

[36]  Renzhi Han,et al.  CRISPR-mediated Genome Editing Restores Dystrophin Expression and Function in mdx Mice. , 2016, Molecular therapy : the journal of the American Society of Gene Therapy.

[37]  J. Joung,et al.  Defining and improving the genome-wide specificities of CRISPR–Cas9 nucleases , 2016, Nature Reviews Genetics.

[38]  Israel Steinfeld,et al.  Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells , 2015, Nature Biotechnology.

[39]  Lynne,et al.  Site-specific integration by adeno-associated virus. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[40]  R. David Hawkins,et al.  Muscle-specific CRISPR/Cas9 dystrophin gene editing ameliorates pathophysiology in a mouse model for Duchenne muscular dystrophy , 2017, Nature Communications.

[41]  Yi-Wei Lee,et al.  In Vivo Delivery of CRISPR/Cas9 for Therapeutic Gene Editing: Progress and Challenges. , 2017, Bioconjugate chemistry.

[42]  Leaf Huang,et al.  Stealth nanoparticles: high density but sheddable PEG is a key for tumor targeting. , 2010, Journal of controlled release : official journal of the Controlled Release Society.

[43]  Feng Zhang,et al.  A split-Cas9 architecture for inducible genome editing and transcription modulation , 2015, Nature Biotechnology.

[44]  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.

[45]  Andrew D. Miller,et al.  pH-triggered nanoparticle mediated delivery of siRNA to liver cells in vitro and in vivo. , 2013, Bioconjugate chemistry.

[46]  M. Gonçalves,et al.  Genome editing at the crossroads of delivery, specificity, and fidelity. , 2015, Trends in biotechnology.

[47]  B. Ghebremedhin.  Human adenovirus: Viral pathogen with increasing importance. , 2014, European journal of microbiology & immunology.

[48]  Xiao-Hui Zhang,et al.  Off-target Effects in CRISPR/Cas9-mediated Genome Engineering , 2015, Molecular therapy. Nucleic acids.

[49]  Jin-Soo Kim,et al.  Analysis of off-target effects of CRISPR/Cas-derived RNA-guided endonucleases and nickases , 2014, Genome research.

[50]  Natalia N. Ivanova,et al.  Increasing the efficiency of homology-directed repair for CRISPR-Cas9-induced precise gene editing in mammalian cells , 2015, Nature Biotechnology.

[51]  H. Kim,et al.  A guide to genome engineering with programmable nucleases , 2014, Nature Reviews Genetics.

[52]  Mark A. Kay,et al.  Progress and problems with the use of viral vectors for gene therapy , 2003, Nature Reviews Genetics.

[53]  Xin Xu,et al.  Nonviral gene editing via CRISPR/Cas9 delivery by membrane-disruptive and endosomolytic helical polypeptide , 2018, Proceedings of the National Academy of Sciences.

[54]  E. Kmiec,et al.  To cleave or not to cleave: therapeutic gene editing with and without programmable nucleases , 2017, Nature Reviews Drug Discovery.

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

[56]  Won Gu Lee,et al.  Electroporation for nanomedicine: a review. , 2017, Journal of materials chemistry. B.

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

[58]  W. Harrington,et al.  A Single Administration of CRISPR/Cas9 Lipid Nanoparticles Achieves Robust and Persistent In Vivo Genome Editing. , 2018, Cell reports.

[59]  Hao Yin,et al.  CRISPR-mediated direct mutation of cancer genes in the mouse liver , 2014, Nature.

[60]  Yi-Wei Lee,et al.  Direct Cytosolic Delivery of CRISPR/Cas9-Ribonucleoprotein for Efficient Gene Editing. , 2017, ACS nano.

[61]  Daniel G. Anderson,et al.  Structure-guided chemical modification of guide RNA enables potent non-viral in vivo genome editing , 2017, Nature Biotechnology.

[62]  Florian Schmidt,et al.  CRISPR genome engineering and viral gene delivery: A case of mutual attraction , 2015, Biotechnology journal.

[63]  Daniel W. Pack,et al.  Design and development of polymers for gene delivery , 2005, Nature Reviews Drug Discovery.

[64]  N. Bessis,et al.  Immune responses to gene therapy vectors: influence on vector function and effector mechanisms , 2004, Gene therapy.

[65]  Denise Serra,et al.  CRISPR/Cas9 Engineering of Adult Mouse Liver Demonstrates That the Dnajb1-Prkaca Gene Fusion Is Sufficient to Induce Tumors Resembling Fibrolamellar Hepatocellular Carcinoma. , 2017, Gastroenterology.

[66]  A. Ehrhardt,et al.  Persistence of high-capacity adenoviral vectors as replication-defective monomeric genomes in vitro and in murine liver. , 2009, Human gene therapy.

[67]  Ning Wang,et al.  Artificial Virus Delivers CRISPR-Cas9 System for Genome Editing of Cells in Mice. , 2017, ACS nano.

[68]  B. Fehse,et al.  Mutagenesis and oncogenesis by chromosomal insertion of gene transfer vectors. , 2006, Human gene therapy.

[69]  R. Lister,et al.  Synthetically controlling dendrimer flexibility improves delivery of large plasmid DNA , 2017, Chemical science.

[70]  C. Leborgne,et al.  Prevalence of serum IgG and neutralizing factors against adeno-associated virus (AAV) types 1, 2, 5, 6, 8, and 9 in the healthy population: implications for gene therapy using AAV vectors. , 2010, Human gene therapy.

[71]  M. Gonçalves,et al.  Engineered Viruses as Genome Editing Devices , 2015, Molecular therapy : the journal of the American Society of Gene Therapy.

[72]  George M. Church,et al.  In vivo gene editing in dystrophic mouse muscle and muscle stem cells , 2016, Science.

[73]  J. Schlehofer,et al.  Update on the prevalence of serum antibodies (IgG and IgM) to adeno‐associated virus (AAV) , 1999, Journal of medical virology.

[74]  Robert Brenner,et al.  Nanoparticle delivery of CRISPR into the brain rescues a mouse model of fragile X syndrome from exaggerated repetitive behaviours , 2018, Nature Biomedical Engineering.

[75]  Atul Kolate,et al.  PEG - a versatile conjugating ligand for drugs and drug delivery systems. , 2014, Journal of controlled release : official journal of the Controlled Release Society.

[76]  G. Ronzitti,et al.  Emerging Issues in AAV-Mediated In Vivo Gene Therapy , 2017, Molecular therapy. Methods & clinical development.

[77]  J. Keith Joung,et al.  High frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells , 2013, Nature Biotechnology.

[78]  Ding Ding,et al.  Quantifying the Ligand-Coated Nanoparticle Delivery to Cancer Cells in Solid Tumors. , 2018, ACS nano.

[79]  Eli J. Fine,et al.  DNA targeting specificity of RNA-guided Cas9 nucleases , 2013, Nature Biotechnology.