RNA Interference Screening Trends 2014
|出版日期||內容資訊||英文 53 Pages
This market report summarizes the results of HTStec's industry-wide global web-based benchmarking survey on RNA interference (RNAi) screening and gene silencing carried out in October 2014.
The survey was initiated by HTStec as part of its tracking of emerging life science marketplaces and to update our previous report in siRNA screening (published September 2012).
The questionnaire was compiled to meet the needs, requirements and interests of the RNAi screening vendor community. The main objectives were to comprehensively document current practices and preferences in RNAi screening, and to understand future user requirements.
Equal emphasis was given to soliciting opinion from all areas and organisations were RNA screening and gene silencing were being undertaken.
The survey looked at RNAi screening as practiced today (2014) and in some cases as predicted for the future (2016). Specifically the following were investigated: key RNAi screening end-user groupings; key diseases or research area(s) targeting RNAi screening; main application areas of RNAi library screening; preferred approach to RNAi screening; types of RNAi screens investigated; main endpoints (readouts) monitored when undertaking RNAi screening; number of different RNAi screening projects attempted per year; level of throughput in RNAi screening projects; use of different cell types in RNAi research; number of different cell lines investigated when attempting RNAi screening; use of different siRNA delivery/transfection methods; how long after transfection are cells typically analyzed; total siRNA concentration typically used in transfections; final assay volume and main plate format for transfection; source of the majority of the siRNAs/shRNAs that make-up respondent's screening library; vendor compiled RNAi library groupings of greatest interest; species against which the majority of siRNAs/shRNAs are directed; size of libraries typically investigated in siRNA/shRNA screens; number of different gene targets silenced; total number of different siRNAs/shRNA constructs in libraries; are siRNAs for one given gene tested individually or as pools; number of siRNAs per pool; main limitations in RNAi library screening; main hurdles in siRNA sample preparation and liquid handling; extent to which automation has been applied to siRNA library screening; most important benefits of applying automation to siRNA screening; main suppliers of siRNA screening instrumentation platforms (i.e. liquid handling, sample prep and automated assay platforms); RNAi screening reagents and consumables budget and its breakdown into components purchased; vendor that first comes to mind when thinking of RNAi screening reagents and consumables; main supplier of RNAi screening reagents and consumables; average cost per single target gene screened by RNAi approaches; aspects of gene silencing where RNAi versus CRISPR approaches are most suited: and any unmet needs that exist in RNAi screening today.
The main questionnaire consisted of 29 multi-choice questions and 1 open-ended question. In addition, there were 6 questions related solely to survey demographics.
The survey collected 50 validated responses, of these 72% provided comprehensive input.
Survey responses were geographically split: 50% Europe; 26% North America; 10% Asia (excluding Japan & China); 8% Japan; 4% Rest of World; and 2% China.
Survey respondents were drawn from persons or groups interested in the gene silencing or RNAi area, and undertaking or planning future investigation of siRNA/shRNA screening.
Respondents came from 28 University/Research Institute/Government Lab/Not-for-Profit Facilities; 6 Large Pharma; 4 Biotech Companies; 3 Medium-Small Pharma; 2 Academic Screening Centers; 2 Contract Research Organizations; 2 Agrochemical/Agri-Biotech Companies; 2 Medical Schools/Hospitals/Clinics; and 1 Biopharma.
Most survey respondents had a senior job role or position which was in descending order: 10 principal investigators; 8 senior scientists/researchers; 6 section/group leaders; 6 directors; 4 professors/assistant professors; 3 research scientists/associates; 3 graduate students; 3 post-docs; 3 department heads; 2 others; 1 lab manager; and 1 vice president.
Survey results were expressed as an average of all survey respondents. In addition, where appropriate the data was fully reanalyzed after sub-division into the following 5 survey groups: 1) HTS/Phenotypic Assays; 2) Basic/Academic Research; 3) Gene/Target ID/Validation; 4) Europe; and 5) North America.
The majority of respondents were currently undertaking siRNA screening, the remainder were not undertaking routinely today, but planned future investigation.
Respondents represented the following key siRNA screening end user groupings: high throughput screening/phenotypic assays (36%); basic/academic research (34%); gene/target identification/validation (22%); clinical research (4%); and other industry/applied research (4%).
Oncology/cancer was the key disease or research area most targeted by RNAi screening.
Identification of new therapeutic targets was the main application area of RNAi screening.
The preferred approach to RNAi screening was primarily an arrayed approach.
The main type of RNAi screens investigated were loss of function screens - based on decrease or loss of a phenotype.
The main endpoints (readouts) monitored when undertaking RNAi screening were either plate-reader based monitoring of whole cell responses or high content microscopic imaging of phenotypic markers/cellular parameters.
A median of 2 to 3 different RNAi screening projects were reported undertaken today (2014).
The RNAi screening throughput achieved today was a median 100-500 wells per week.
The cell type most used today for RNAi research and screening was tumor cell lines.
A median of 2 to 3 different cell lines were investigated per gene silencing project today.
The siRNA delivery method most used today was standard chemical (lipid-mediated) transfection.
Cells were analysed for silencing a median of 72 hours after siRNA transfection.
The median total siRNA concentration used in transfection today was 10-30nM.
The final assay volume used for siRNA transfection today was a median of 25µL-50µL.
The microplate format most used for siRNA delivery (transfection) and screening today was 384-well (standard volume).
The majority of siRNAs/shRNA constructs that make up respondent's primary screening library were sourced as off-the-shelf purchases of individual siRNAs.
The vendor compiled RNAi libraries of most interest to respondents covered the entire Human Genome.
The species that the majority of siRNAs/shRNAs in respondent's libraries were directed against was Human.
The median library size investigated in siRNAs/shRNAs screens were: 500-1K gene targets silenced; representing 100-500 different siRNAs/shRNAs; nearly half of siRNAs were tested as pools with a median of 4 siRNAs per pool.
Off-target effects were ranked as what most limits work on RNAi screening today.
Optimising transfection conditions and minimising cytotoxicity were rated as the aspect of siRNA sample prep and liquid handling that most limits RNAi screening efficiency.
Only moderate to low levels of automation have been most applied to siRNA screening today.
Enhanced and more uniform transfection efficiency was rated the most important potential benefit of applying automation to siRNA screening.
The suppliers of siRNA screening instrumentation most used by respondents came mainly from Thermo Scientific/Life Technologies, PerlinElmer, Tecan and BioTek.
The median annual budget for RNAi screening reagents and consumables last year (2013) was $25K-$50K per lab per year.
A bottom-up model was developed around respondent's annual budget to estimate the global market for RNA screening reagents and consumables. In 2013 this market was estimated to be around $40M. Segmentation & CAGR estimates are given in the full report.
The breakdown of the annual RNAi screening reagents and consumables budget saw the greatest share allocated to transfection reagents and buffers.
The supplier/vendor of RNAi screening reagents and consumables that first comes into the mind of respondents was Thermo Scientific/Life Technologies.
The suppliers/vendors of RNAi screening reagents and consumables most used by respondents were Thermo Scientific/Life Technologies and GE Dharmacon.
The median cost per single target gene investigated by RNAi screening approaches was $1-$2.5.
RNAi approaches were preferred over CRISPR for most technology attributes related to gene silencing.
All Respondents feedback on any unmet needs that exist today in RNAi screening were documented.
The full report provides the data, details of the breakdown of the responses for each question, its segmentation and the estimates for the future (2016). It also highlights some interesting differences between survey groups.