Cas9 protein delivery non-integrating lentiviral vectors for gene correction in sickle cell disease.
Naoya Uchida,Naoya Uchida,Claire M. Drysdale,Tina Nassehi,Jackson Gamer,Morgan Yapundich,Julia DiNicola,Yoshitaka Shibata,Malikiya Hinds,Bjorg Gudmundsdottir,Juan J. Haro-Mora,Selami Demirci,John F. Tisdale +12 more
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TL;DR: In this article, a Cas9 protein delivery non-integrating lentiviral system encoding guide RNA and donor DNA, allowing for transient endonuclease function and inclusion of all editing tools in a single vector (all-in-one).
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Abstract: Gene editing with the CRISPR-Cas9 system could revolutionize hematopoietic stem cell (HSC)-targeted gene therapy for hereditary diseases, including sickle cell disease (SCD). Conventional delivery of editing tools by electroporation limits HSC fitness due to its toxicity; therefore, efficient and non-toxic delivery remains crucial. Integrating lentiviral vectors are established for therapeutic gene delivery to engraftable HSCs in gene therapy trials; however, their sustained expression and size limitation preclude their use for CRISPR-Cas9 delivery. Here, we developed a Cas9 protein delivery non-integrating lentiviral system encoding guide RNA and donor DNA, allowing for transient endonuclease function and inclusion of all editing tools in a single vector (all-in-one). We demonstrated efficient one-time correction of the SCD mutation in the endogenous βs-globin gene up to 42% at the protein level (p
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Citations
Delivery of CRISPR-Cas tools for in vivo genome editing therapy: Trends and challenges
TL;DR: In this article , a review summarizes existing in-vivo genome editing delivery systems and possible solutions to overcome their limitations, highlighting the ongoing clinical trials for in vivo genome editing therapy and recently developed genome editing tools for their potential applications.
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Viral Vectors for the in Vivo Delivery of CRISPR Components: Advances and Challenges
TL;DR: This review thoroughly examined recent achievements in using a variety of viral vectors as a means of CRISPR/Cas9 delivery, as well as the benefits and limitations of each viral vector.
Current approaches in CRISPR-Cas9 mediated gene editing for biomedical and therapeutic applications.
Gargi Bhattacharjee,N. M. Gohil,Khushal Khambhati,Indra Mani,Rupesh Maurya,Janardhan Keshav Karapurkar,J H Gohil,Dinh-Toi Chu,Hue Vu Thi,K. Alzahrani,Pau Loke Show,Rakesh Rawal,S. Ramakrishna,Vijai Singh +13 more
TL;DR: In this paper , the authors highlight recent developments and utilization of the CRISPR-Cas9 system in correcting or generating gene mutations to create model organisms to develop deeper insights into diseases, rescue normal gene functionality, and curb the progression of a disease.
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Non-Integrating Lentiviral Vectors in Clinical Applications: A Glance Through
TL;DR: The use of non-integrating LVs (NILVs) reduces insertional mutagenesis and the risk of malignant cell transformation over integrating lentiviral vectors, especially in non-dividing cells.
Immunogenicity of CRISPR therapeutics—Critical considerations for clinical translation
TL;DR: In this paper , the authors examine what is currently known about the immunogenicity of CRISPR therapeutics and discuss several considerations to mitigate immunogenity for the design of safe and clinically translatable CRISpr therapeutics.
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