Crispr Application
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The document outlines various applications of the CRISPR/Cas9 technology in basic and medical research, highlighting its role in gene expression regulation, disease modeling, and drug discovery. It emphasizes the ability of CRISPR to create accurate animal and cell models for studying diseases, facilitate the development of therapies, and enhance drug screening processes. Additionally, it discusses innovative techniques such as CRISPR-mediated imaging and RNA editing, reflecting on the future potential of gene editing in biotechnology.
Crispr Application
- 1. CRISPR/Cas9 Applications Tel: 1-631-626-9181 Fax: 1-631-614-7828 Email: info@creative-biogene.com https://www.creative-biogene.com/crispr-cas9/
- 2. 1 Basic research 2Medical research 3 Biotechnology development CONTENTS powerful new tools in laboratory research promise approach to human disease treatment drive innovative applications to biotechnology
- 3. CRISPR/Cas9 in Basic Research 1 G e n e E x p r e s s i o n a n d E p i g e n e t i c R e g u l a t i o n A n i m a l / C e l l M o d e l s Develop animal/cell models to promote understanding of gene functions and even disease processes. Transcriptional regulation Epigenetic modification
- 4. 01 Gene conventional knock out by NHEJ 03Knockin/Point mutation 02 Conditional knockout/knockin 04Chromosome fusion Animal/Cell Models 1.1 The CRISPR/Cas system simplifies the functional study of genes in cells and animals (such as yeast, fish, mice and many other animals) in an unprecedented way; Develop more accurate models (especially hereditary diseases, neurodegenerative diseases, cardiovascular diseases, and cancer) to promote understanding of human disease processes and to provide directions for developing treatments to address these deficiencies.
- 5. Epigenetic modification Using non-active dCas9 (another type of Cas9 lacking nuclease activity but retaining DNA binding activity) fused enzymes such as DNA methylase, histone acetyltransferase, and deacetylase can be targeted to alter the epigenetic state of precise locations within the genome. Gene Expression and Epigenetic Regulation 1.2
- 6. CRISPR-mediated transcriptional modulation dCas9 can also be fused to transcriptional repressors or activators targeting the promoter region, and these dCas9 fusion proteins can lead to strong transcriptional repression (CRISPR interference or CRISPRi) or activation (CRISPRa) of downstream target genes. Gene Expression and Epigenetic Regulation 1.2 CRISPRi CRISPRa
- 7. CRISPR/Cas9 in Medical research 02 01 Drug discovery02 Treatment of disease03 Human disease models
- 8. CRISPR/Cas9-mediated human disease models The CRISPR/Cas9 system has been used to generate many disease-based models for many important human diseases, including neurological diseases, cardiovascular diseases and cancer, as well as other Mendelian or complex genetic human diseases, which make it possible to study the molecular mechanisms of the underlying pathogenesis, drug screening and discovery, high-throughput research, and gene therapy. CRISPR/Cas9 can also efficiently target genes in somatic tissues, a method that is particularly useful for studying age-related disease physics. 2.1
- 9. Screening for target sites The CRISPR library can detect living cells with specific conditions, such as drug therapy. By using the system, researchers can identify genes and proteins that cause or prevent disease, thereby identifying potential drug targets. Drug discovery CRISPR/Cas9 animal models allow scientists to discovery new drugs more accurately and verify the safety and efficacy of the drugs, ensuring that these models better predict what will happen in clinical trials. Upregulating or downregulating gene activity using the CRISPR/Cas9 system is a subtle way of studying the importance of genes and proteins that can be activated or inhibited by drugs to treat disease. Drug discovery 2.2
- 10. Well-known pharmaceutical companies and emerging biotech companies are racing to develop CRISPR-based therapies. Compared to other gene therapy strategies, CRISPR genome editing is considered faster, cheaper, and pot entially safer. The CRISPR genome editing is especially useful for diseases that can be resolved by modifying cells. Autologous CRISPR cell therapy is promising in bypassing the exclusion problems that exist in donor-matched transpla ntation therapies. Treatment of disease 2.3
- 11. CRISPR/Cas9-mediated ChIP01 Live Imaging of the Cellular Genome via CRISPR System02 03CRISPR System in Biotechnology Research RNA Editing03
- 12. CRISPR/Cas9-mediated ChIP 3.1 Epitope tags can be fused to dCas9 for efficient purification, including 3xFLAG-tags, biotin tags, etc. The locus is then isolated by affinity purification against the epitope tag. Afte r purification of the locus, the locus-associated molecules can be identified by mass spectr ometry (protein), RNA sequencing (RNA) and NGS (other genomic regions).
- 13. Live Imaging of the Cellular Genome via CRISPR System 3.2 One method uses orthogonal dCas9 labeled with different fluorescent proteins. Another method gRNA that interacts with orthologous proteins that recruit specific orthogonal RBPs labeled with different fluorescent proteins. 01 02 CRISPR imaging provides a unique method to detect chromatin dynamics in living cells and has many advantages, including the simplicity of gRNA design, ease of implementation, programmable for different genomic loci, the ability to detect multiple genomic loci, and compatibility with live-cell imaging.
- 14. RNA Editing with CRISPR/Cas13 3.3 CRISPR/Cas13 provides a transient gene expression suppression tool in mammalian cells RNA-targeting CRISPR/Cas13 can be used for RNA editing as a catalytically dead Cas13 (dCas13) variant fused to the deaminase domain of human ADAR (ADARDD). 01 02 dCas13b can be coupled to fluorescent proteins (e.g. EGFP) to tracking the translocation of endogenous transcripts from the nucleus to the cytoplasm. In addition, dCas13 can be fused to functional effector domains for posttranscriptional manipulations of RNA substrates in living mammalian cells (e.g. splicing, RNA translation, and others). 03
Editor's Notes
- #5: The rapid development of the versatility of the CRISPR/Cas system opens up many ways to manipulate gene expression, which have simplified the functional study of genes in cells and experimental animals such as yeast, fish, mice and many other animals in an unprecedented way. Developed more accurate models (especially genetic diseases, neurodegenerative diseases, cardiovascular diseases, and cancer) to promote understanding of human disease processes, and also provided directions for developing treatments to counter these defects.