Genome Editing Review - Recent Technologies & Potential Applications
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The document provides an overview of gene editing technologies, highlighting their history, types, and applications, particularly focusing on CRISPR technology. It discusses the rapid growth predicted for the gene editing market, driven by advancements such as CRISPR and increasing investments in research for healthcare and agriculture. Additionally, the document outlines the distinctions between forward and reverse genetics and various gene editing methods, emphasizing their potential in clinical therapies, cancer research, and food sustainability.
Genome Editing Review - Recent Technologies & Potential Applications
- 1. Recent Technologies & Potential Applications For more info, contact us: xeraya@xeraya.com Follow us: @xerayacapital www.xeraya.com Genome Editing Review June 2022. © Xeraya Capital. 1
- 2. Genome Editing Review Content Overview • Recap: What is Gene Editing? • Brief History • Forward vs Reverse Genetics • The 3 Distinct Terms • Outcomes: Homologous vs non-Homologous • Genome Editing Technologies • Applications of Gene Editing Technology June 2022. © Xeraya Capital. 2
- 3. Recap: What is Gene Editing? Gene editing (or genome editing) is a group of technologies that give scientists the ability to change an organism's DNA. These technologies allow genetic material to be added, removed, or altered at specific locations in the genome. June 2022. © Xeraya Capital. 3 Credit: iStock Photos Source: https://medlineplus.gov/genetics/understanding/genomicresearch/genomeediting/
- 4. A Brief History of Gene Editing June 2022. © Xeraya Capital. 4 Source: https://geneeditinginstitute.com/about/our-history/
- 5. Key Figures in Gene Editing June 2022. © Xeraya Capital. Gene Editing Market projected at USD 19.4 billion in 2028 - Increase from USD 5.2 billion in 2020 - CAGR of 22.9% (2021-2028) - Increased use of CRISPR tech. against coronavirus Grand View Research USD 5.1 billion in 2021 Big pharma & biotech invest in gene editing tech. for COVID-19 vaccine R&D. CRISPR has largest revenue share Market share of 48.8% in 2020, revenue up to USD 1.5 billion (followed by ZFNs at USD 1.3 billion). Source: Grand View Research, Global Market Insights & MarketsAndMarkets 5 Key Drivers in Gene Editing: - R&D to combat COVID-19 (from diagnostic to kill- sequence) - Increased prevalence of chronic & genetic disorders - Rising government support in gene R&D - Growing demand for synthetic genes Global Market Insights
- 6. Terms & Concepts Understanding Gene Editing at its Core June 2022. © Xeraya Capital. 6
- 7. Forward Genetics vs Reverse Genetics June 2022. © Xeraya Capital. 7 Phenotype ‣ Genotype Forward genetics identifies & links a mutation to a disease. Altered or abnormal phenotype examined by screening genes. Genotype ‣ Phenotype Reverse genetics induces mutations in model organisms to study their role in disease. The genotype is altered. FORWARD REVERSE Source: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5002245/ Phenotype: observable physical properties i.e., appearance, development, behavior. Genotype: the genetic markup of an organism, describes an organism’s complete set of genes.
- 8. Three Distinct Terms June 2022. © Xeraya Capital. 8 Source: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7596157/ Genome Engineering Sequence of genomic DNA is designed and modified Genome Editing Site-specific modifications into genomic DNA using DNA repair mechanisms Gene Editing Modification of a single gene (enough DNA to code for one protein)
- 9. Major Mechanisms of Gene Editing June 2022. © Xeraya Capital. 9 Source: https://www.nature.com/articles/s41392-019-0089-y Nuclease-induced DNA double strand breaks (DSBs) activates one of two major repair mechanisms that occur in almost all cell types: • nonhomologous end- joining (NHEJ) that is error prone, OR • homology-directed repair (HDR) that is more precise
- 10. Gene Editing Technologies Overview of MegNs, ZFNs, TALENs & CRISPR/Cas June 2022. © Xeraya Capital. 10
- 11. Meganucleases (MegNs) Endodeoxyribonucleases with large recognition site (DNA sequences of 12-40 base pairs); making the site generally occurring only once in any given genome. Given its high specificity, the probability of finding an enzyme that targets a desired locus is small and production of customized MegNs remains complex & highly inefficient. June 2022. © Xeraya Capital. 11 Credit: Castro et al., IJMS Source: https://en.wikipedia.org/wiki/Meganuclease First identified in the 1990s, researchers discover that applications of MegNs in genome editing is limited and technically challenging to work with.
- 12. Zinc finger Nucleases (ZFNs) Artificially engineered restriction enzymes for custom site-specific genome editing. ZFNs themselves are transcription factors, where each finger recognizes 3-4 bases. While ZFNs are easier to design compared to MegNs, its prone to cause off-target cleavages that leads to apparent toxicity to cells. Source: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7596157/ Credit: Papaioannou, et al., ResearchGate June 2022. © Xeraya Capital. 12
- 13. Transcription Activator-Like Effector nucleases (TALENs) TALENs are similar to ZFNs and MegNs in that the proteins must be re-engineered for each targeted DNA sequence. It is however, complicated for whom not familiar with molecular biological experiments. It is also confronted with some limitations, such as their large size (impeding delivery) in comparison to ZFNs. June 2022. © Xeraya Capital. 13 Compared ZFNs, a single TALEN module can recognize one nucleotide, making them simpler to design with lower number of off-target breaks. Source: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7596157/
- 14. CRISPR/Cas system Short for clustered regularly interspaced short palindromic repeats, the CRISPR/Cas system is the most recent (and most popular) platform in the field of genome editing. It is low cost, easy to design and highly scalable. Unlike ZFNs & TALENs, the DNA sequence recognition of the CRISPR/Cas system is based on RNA-DNA interactions. Source: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7596157/ Credit: StockAdobe.com June 2022. © Xeraya Capital. 14 The CRISPR/Cas system was developed in 2013 and is known as the third-generation genomic editing tool.
- 15. Gene editing Platforms Compared June 2022. © Xeraya Capital. 15 Source: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7596157/ Feasibility MegNs (obsolete) ZFNs TALENs CRISPR/Cas9 Design Simplicity --- - + ++ Cost & affordability -- + - ++ Scalability -- - -/+ + Site recognition length (bp) 12-40 18-36 28-40 Up to 44 Targeting & cleavage efficacy -/+ - + ++ Programmability --- -- - + Success Rate (est.) n/a 24% 99% 90%
- 16. Gene Editing Applications From Clinical Research & Therapy to Food Sustainability June 2022. © Xeraya Capital. 16
- 17. Various Applications of Gene Editing June 2022. © Xeraya Capital. 17 Crops Viral Diseases Gene Therapy Cancer Research Rare Diseases Source: https://www.nature.com/articles/s41392-019-0089-y
- 18. Gene Editing in Clinical Therapy June 2022. © Xeraya Capital. 18 Source: https://www.nature.com/articles/s41392-019-0089-y Ex vivo: Cells are isolated from a patient to be treated, edited and re-engrafted back to the patient. For therapeutic success, the target cells must survive in vitro and return to the target tissue after transplantation. In vivo: Engineered nucleases are delivered (viral or non-viral) and directly injected into the patient for systemic or targeted tissue (such as the eye, brain, or muscle) effect. Credit: Li, et al., Nature.com (2020)
- 19. Gene Editing in Cancer Immunotherapy June 2022. © Xeraya Capital. 19 One promising area in immunotherapy is the application of genetically engineered T cells, known as chimeric antigen receptor (CAR) T cells, which allow the targeting of tumor- associated antigens and could enhance the therapy response. Using CRISPR/Cas9, Liu and his colleagues efficiently generated CAR T cells in which 2 or 3 genes were simultaneously disrupted and tested their antitumor function both in vitro and in vivo. Source: https://www.nature.com/articles/s41392-019-0089-y Credit: Li, et al., Nature.com (2020)
- 20. Using CRISPR for food sustainability The agriculture industry needs to solve an extremely complex problem — feeding more people while facing increasingly dire resource constraints. While conventional plant breeding relies on introducing random changes into plant DNA, CRISPR technology offers precise alterations to specific DNA sequences. Source: https://www.genengnews.com/topics/genome-editing/from- pharma-to-farm-can-crispr-feed-the-world/ June 2022. © Xeraya Capital. 20 Inari focuses on soy, corn & wheat – major crops that feed the world & impact the environment the most. TreeCo is using CRISPR technology to enhance trees so it can be bred about 10x faster
- 21. • Genome editing is a fast-growing field. Much progress has been accomplished in the improvement of gene editing technologies since their discovery. • The tremendous advances in the development of engineered nucleases (especially ZFNs, TALENs, and CRISPR/Cas9) paved the way for genome editing from a theoretical concept into clinical practice. • CRISPR/Cas9 system has been increasingly popular in recent years, due its relatively low cost. Researchers found that it is easy to design, highly scalable and has a relatively high success rate. • Gene editing applications include manufacturing engineered medical products, eradication of human genetic diseases, treatment of AIDS and cancers, as well as improvement of crop and food in coming years. Conclusion June 2022. © Xeraya Capital. 21
- 22. By xeraya capital For more info, contact us: xeraya@xeraya.com Follow us: @xerayacapital www.xeraya.com June 2022. © Xeraya Capital. 22