from the evidence figure i gave you is there any figure that shows crispr fixing single gene disorder
The median time period since survey respondents had first investigated or started using CRISPR/Cas 9 gene editing technology was 6-12 months ago (Figure 2).Most (47%) of survey respondents were applying or intending to apply CRISPR/Cas 9 gene editing to the basic research area of the drug discovery process. This was followed by target validation (18% applying); target identification (10% applying) and then clinical studies and lead generation (both with 9% applying).
Figure 2 Time period since first started using CRISPR/Cas9 gene editing technology
Other drug discovery areas had in total only 6% applying (Figure 3).
Figure 3 Area of the drug discovery process most applying or intending to apply CRISPR/Cas9 gene editing
Oncology/cancer was the key disease or research area most targeted by survey respondents (52% targeting) by CRISPR/Cas 9 gene editing. This was followed by immunology/inflammatory disease/ autoimmune (30% targeting); neurology/CNS/ neurodegeneration/pain (27% targeting); metabolic disease/diabetes (18% targeting); and then cardiovascular disease (17% targeting) (Figure 4).
Figure 4 Key diseases or research areas using CRISPR/Cas9 gene editing
Most (49%) of survey respondents answered N/A, ie they have not investigated other gene editing technologies prior to CRISPR/Cas 9 availability. 29% have previously investigated transcription activator-like effector nucleases (TALENs); 21% have investigated integration via homologous recombination (eg with rAAV); 16% zinc finger nucleases (ZFN); and 11% other approaches (Figure 5).
Figure 5 Other gene editing technologies investigated prior to CRISPR/Cas9
Main rationale for using CRISPR/Cas9 gene editing technology
A gene knockout was what the majority (77%) of survey respondents most want to achieve using CRISPR/Cas 9 gene editing technology. This was followed by introduce defined mutations, insertions or modifications (62% wanting); gene knock-in (52% wanting); and then gene knock down (inducible) (40% wanting). Other aims were wanted by less than a third of survey respondents (Figure 6).
Figure 6 What respondents want to achieve using CRISPR/Cas9 gene editing
Identification of new therapeutic targets was the main objective of survey respondents CRISPR/Cas 9 gene editing in drug discovery (61% wanting). This was followed by validation of new therapeutic targets (48% wanting); and then investigation of mechanism of action and screens to identify genes that regulate various cell biological processes (both with 46% wanting). Of least interest was deconvolution and validation of GWAS hits (only 8% targeting) (Figure 7).
Figure 7 Main objectives of CRISPR/Cas9 gene editing in drug discovery
Survey respondents ranked some of the perceived advantages of CRISPR/Cas 9 gene editing technology in order of importance. This analysis revealed that efficient, ie edit targets sequences at surprisingly high rates, was seen as the main advantage of CRISPR/Cas 9 gene editing technology. This was followed by simplicity, ie easy to use and design; programmable, ie get precision targeting; and then fast, ie get tangible results within weeks. Ranked least advantageous was multiplexing, ie can programme multiple guide RNAs and cleave multiple genes simultaneously (Figure 8).
Figure 8 The advantages of CRISPR/Cas9 gene editing technology
Survey respondents rated complete genetic knockout, while minimising off-target effects as the potential benefit of CRISPR/Cas 9 in drug discovery they were most interested in exploiting. This was closely followed by enables rapid generation of cell lines harbouring desired mutations; develop accurate models of complex human disease in an efficient manner; and then relative ease with which cellular models can be generated. Rated least interesting was scalable generation of genome-wide CRISPR libraries for HT functional genomics screening (Figure 9).
Figure 9 Potential benefits of CRISPR/Cas9 in drug discovery most respondents want to exploit
Validating gene edits
The downstream analytical technique most used to validate CRISPR/Cas 9 gene edits was PCR (70% using). This was followed by real time PCR (59% using); Western blotting (55% using); FACS (34% using) and then next gen sequencing (miSEQ) (30% using). Least used were capillary electrophoresis, TIDE assay and other DNA mismatch assays (Figure 10).
Figure 10 Downstream analytical techniques used to validate CRISPR/Cas9 gene editing
Multiplexing gene edits
For each single target gene survey respondents have attempted to edit they have designed a median of two single guide RNAs (ie 2 sgRNAs). Interest in simultaneously delivering sgRNAs targeting multiple genes to multiplex CRISPR/Cas 9 gene edits is presented in Figure 11. This showed that the majority (69%) of survey respondents are not yet undertaking multiplexing, but plan future investigation. This was followed by 25% not undertaking with no plans to investigate, leaving only 5% actually undertaking multiplexing today (Figure 11).
1 answer