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Title: Delivering CRISPR: a review of the challenges and approaches

Gene therapy has long held promise to correct a variety of human diseases and defects. Discovery of the Clustered Regularly-Interspaced Short Palindromic Repeats (CRISPR), the mechanism of the CRISPR-based prokaryotic adaptive immune system (CRISPR-associated system, Cas), and its repurposing into a potent gene editing tool has revolutionized the field of molecular biology and generated excitement for new and improved gene therapies. Additionally, the simplicity and flexibility of the CRISPR/Cas9 site-specific nuclease system has led to its widespread use in many biological research areas including development of model cell lines, discovering mechanisms of disease, identifying disease targets, development of transgene animals and plants, and transcriptional modulation. In this review, we present the brief history and basic mechanisms of the CRISPR/Cas9 system and its predecessors (ZFNs and TALENs), lessons learned from past human gene therapy efforts, and recent modifications of CRISPR/Cas9 to provide functions beyond gene editing. We introduce several factors that influence CRISPR/Cas9 efficacy which must be addressed before effective in vivo human gene therapy can be realized. The focus then turns to the most difficult barrier to potential in vivo use of CRISPR/Cas9, delivery. We detail the various cargos and delivery vehicles reported for CRISPR/Cas9, including physical delivery methods (e.g.more » microinjection; electroporation), viral delivery methods (e.g. adeno-associated virus (AAV); full-sized adenovirus and lentivirus), and non-viral delivery methods (e.g. liposomes; polyplexes; gold particles), and discuss their relative merits. We also examine several technologies that, while not currently reported for CRISPR/Cas9 delivery, appear to have promise in this field. The therapeutic potential of CRISPR/Cas9 is vast and will only increase as the technology and its delivery improves.« less
Authors:
 [1] ;  [1] ;  [1] ; ORCiD logo [1]
  1. Sandia National Lab. (SNL-NM), Albuquerque, NM (United States). Bioenergy and Defense Technologies
Publication Date:
Report Number(s):
SAND-2018-10248J
Journal ID: ISSN 1071-7544; 667983
Grant/Contract Number:
NA0003525
Type:
Accepted Manuscript
Journal Name:
Drug Delivery
Additional Journal Information:
Journal Volume: 25; Journal Issue: 1; Journal ID: ISSN 1071-7544
Publisher:
Taylor & Francis
Research Org:
Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)
Sponsoring Org:
USDOE National Nuclear Security Administration (NNSA); SNL Laboratory Directed Research and Development (LDRD) Program
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; CRISPR; Cas9; history; overview; review; drug delivery; prospective
OSTI Identifier:
1474089

Lino, Christopher A., Harper, Jason C., Carney, James P., and Timlin, Jerilyn A.. Delivering CRISPR: a review of the challenges and approaches. United States: N. p., Web. doi:10.1080/10717544.2018.1474964.
Lino, Christopher A., Harper, Jason C., Carney, James P., & Timlin, Jerilyn A.. Delivering CRISPR: a review of the challenges and approaches. United States. doi:10.1080/10717544.2018.1474964.
Lino, Christopher A., Harper, Jason C., Carney, James P., and Timlin, Jerilyn A.. 2018. "Delivering CRISPR: a review of the challenges and approaches". United States. doi:10.1080/10717544.2018.1474964. https://www.osti.gov/servlets/purl/1474089.
@article{osti_1474089,
title = {Delivering CRISPR: a review of the challenges and approaches},
author = {Lino, Christopher A. and Harper, Jason C. and Carney, James P. and Timlin, Jerilyn A.},
abstractNote = {Gene therapy has long held promise to correct a variety of human diseases and defects. Discovery of the Clustered Regularly-Interspaced Short Palindromic Repeats (CRISPR), the mechanism of the CRISPR-based prokaryotic adaptive immune system (CRISPR-associated system, Cas), and its repurposing into a potent gene editing tool has revolutionized the field of molecular biology and generated excitement for new and improved gene therapies. Additionally, the simplicity and flexibility of the CRISPR/Cas9 site-specific nuclease system has led to its widespread use in many biological research areas including development of model cell lines, discovering mechanisms of disease, identifying disease targets, development of transgene animals and plants, and transcriptional modulation. In this review, we present the brief history and basic mechanisms of the CRISPR/Cas9 system and its predecessors (ZFNs and TALENs), lessons learned from past human gene therapy efforts, and recent modifications of CRISPR/Cas9 to provide functions beyond gene editing. We introduce several factors that influence CRISPR/Cas9 efficacy which must be addressed before effective in vivo human gene therapy can be realized. The focus then turns to the most difficult barrier to potential in vivo use of CRISPR/Cas9, delivery. We detail the various cargos and delivery vehicles reported for CRISPR/Cas9, including physical delivery methods (e.g. microinjection; electroporation), viral delivery methods (e.g. adeno-associated virus (AAV); full-sized adenovirus and lentivirus), and non-viral delivery methods (e.g. liposomes; polyplexes; gold particles), and discuss their relative merits. We also examine several technologies that, while not currently reported for CRISPR/Cas9 delivery, appear to have promise in this field. The therapeutic potential of CRISPR/Cas9 is vast and will only increase as the technology and its delivery improves.},
doi = {10.1080/10717544.2018.1474964},
journal = {Drug Delivery},
number = 1,
volume = 25,
place = {United States},
year = {2018},
month = {5}
}

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