How Genome Editing is Revolutionizing Medicine

Contents

1 Introduction

2 State-of-the-art Gene Editing
2.1 PAM sequence
2.2 Type II CRISPR-Cas System - Cas9 variants and orthologs
2.2.1 dCas9
2.2.2 eSpCas9
2.2.3 SpCas9-HF1
2.2.4 HypaCas
2.2.5 xCas
2.2.6 Cas9 Nickases
2.2.7 Remarkable SpCas9 Orthologs
2.2.8 Base editing
2.2.9 Prime editing
2.3 Type V CRISPR-Cas System - Diagnostic tools
2.3.1 Cas12a, Cas12b
2.4 Type VI CRISPR-Cas System - RNA targeting
2.4.1 Cas13a
2.4.2 CasRx
2.5 Transfection methods
2.5.1 Non-viral
2.5.2 Viral

3 Gene Editing and Treatment of Diseases
3.1 Hemoglobinopathies
3.1.1 CRISPR-correction of sickle cell disease
3.1.2 Reactivation of Fetal Hemoglobin Production
3.2 Cystic fibrosis
3.2.1 Functional repair of the CFTR gene
3.2.2 CFTR repair using Adenine Base Editors
3.3 Duchenne muscular dystrophy (DMD)
3.3.1 CRISPR-Cas9 restores functional dystrophin in mice
3.3.2 Long-term evaluation of Dystrophin restoration in mice
3.4 Treatment of Human Immunodeficiency Virus
3.4.1 Genome editing of HIV co-receptors CCR5 and CXCR4
3.4.2 HIV elimination using ART and CRISPR-Cas9
3.5 Cancer immunotherapy
3.5.1 Editing Primary Human Natural Killer Cells
3.5.2 Creating enhanced CAR T Cells using CRISPR-Cas9
3.5.3 Creating universal CAR T cells with CRISPR-Cas9

4 Gene Editing for Drug Development
4.1 Glycoengineering
4.2 Prolonged cultivation
4.3 Elimination of host cell protein impurities
4.4 Rapid adaptation cells
4.5 Enhanced protein production

5 Gene Editing Tools for Diagnostics
5.1 Diagnostic CRISPR endonucleases
5.1.1 Cas12a, Cas12b
5.1.2 Advanced endonucleases for diagnostic
5.1.3.. Cas13a
5.2 SHERLOCK
5.2.1 SARS-CoV-2 Detection using SHERLOCK
5.3 DETECTR 29
5.3.1 SARS-CoV-2 Detection using DETECTR
5.4 CRISPR targeting of drug-resistant bacteria
5.4.1 Targeting pathogenic and drug-resistant bacteria

6 Challenges and Obstacles
6.1 Off-target effects and carcinogenesis
6.2 Delivery of the CRISPR-Cas system
6.3 Immune responses against CRISPR-Cas

7 Prospect

List of Figures

List of Abbreviations

Bibliography