Convenient rapid and portable tool for the detection of ribonucleases

Document text

Principal Investigator: LANCE P FORD
Organization: ATTOGENE
Fiscal Year: 2023
Award: $229,975
Funding agency: National Institute of General Medical Sciences

Summary/Abstract
 The past decade has brought an escalating growth of important new commercial applications for RNA.
RNA molecules have emerged as the preeminent regulators of gene expression. They have a unique ability to
edit genomes and destabilize gene products. New diagnostic applications for RNA are continually being
discovered. Last but not least, modified mRNA nanoparticle formulations are enabling the development of new
vaccines and cellular therapies with a speed and efficacy unrivaled by other processes. This newfound
success has driven increased interest and efforts in many areas of RNA research. Researchers are constantly
discovering important new roles for RNA in the regulation of cellular processes. It has also opened the door to
many new commercial opportunities for startups and large pharmaceutical and diagnostic companies alike.
Accordingly, massive effort is now directed to production scale-up and large-scale manufacturing of RNA
molecules. As production increases, there is a far greater need for care and control of manufacturing
processes. This is especially true for RNA since it is quite vulnerable to degradation by contaminating enzymes
in the surrounding environment.
 As production scales and costs increase, more attention must be paid to strict process control and
quality management. For example, manufacturing of RNA for regulated therapeutic and diagnostic applications
must be performed in a certified RNAse-free environment. Current commercial methods to verify the absence
of RNAse require expensive equipment and hours to perform. The cost and time required to perform RNAse
testing hinders the productivity of manufacturers and researchers alike. A new rapid tool to test for unwanted
RNA contamination would great assist researchers and manufacturers who must frequently test their
environments and supply chain materials.
 In Phase I, we will create a novel prototype lateral flow test for the sensitive detection of RNAse. Our
work will produce a test kit product with several important advantages over currently available tools:
Convenience: Our test will be portable and not require bulky, expensive lab equipment. Sensitivity: Our test will
be at least 5-times more sensitive than current methods. Time: Users will see their results in less than 15
minutes rather than hours. These combined advantages of our proposed product will create a highly effective
tool to further support the continuing innovation in RNA therapeutics and research.
Specific Aim #1: Optimize the incubation buffer and RNA oligonucleotide sequence to improve RNAse assay
sensitivity.
Specific Aim #2: Produce a more sensitive fluorescent version of the lateral flow RNAse detection assay.
Specific Aim #3: Enable testing of solid surfaces using swab wipes.

Terms: <Accidents><Address><Area><Assay><Attention><Basic Research><Basic Science><Binding><Bioassay><Biologic Assays><Biological Assay><Biology><Buffers><COVID-19><COVID19><CV-19><CV19><Caring><Catalysis><Cell Function><Cell Process><Cell Therapy><Cell physiology><Cellular Function><Cellular Physiology><Cellular Process><Cellular Regulation><Certification><Communities><DNA><Data><Data Analyses><Data Analysis><Deoxyribonucleic Acid><Detection><Detergents><Development><Devices><Diagnostic><Diagnostic Reagent Kits><Disease><Disorder><Environment><Enzyme Gene><Enzymes><Equipment><Fluorescence><Formulation><Freeze Drying><Freeze Dryings><Gene Expression><Generalized Growth><Goals><Growth><High temperature of physical object><Hour><Incubated><Industry><Investigators><Lateral><Liquid substance><Lyophilization><Manufacturer><Messenger RNA><Methods><Modification><Molecular Interaction><Monitor><Non-Polyadenylated RNA><Oligo><Oligonucleotides><Pancreatic RNase><Pancreatic ribonuclease><Pharmaceutical Agent><Pharmaceuticals><Pharmacologic Substance><Pharmacological Substance><Phase><Procedures><Process><Production><Productivity><Q-Dot><Quantum Dots><RNA><RNA Gene Probes><RNA Gene Products><RNA Nucleases><RNA Probes><RNA based therapeutics><RNA based therapy><RNA therapy><RNA vaccine><RNA-based vaccine><RNase><RNase A><RNase I><Reagent><Regulator Genes><Research><Research Personnel><Researchers><Resistance><Ribonuclease A><Ribonuclease Family Protein><Ribonuclease I><Ribonucleases><Ribonucleic Acid><Role><Sampling><Sodium Chloride><Solid><Speed><Strepavidin><Streptavidin><Subcellular Process><Surface><Swab><System><Testing><Therapeutic><Time><Tissue Growth><Transcriptional Regulatory Elements><Tube><VAC-TX><Vaccine Production><Vaccine Therapy><Vaccines><Visualization><Work><cell growth regulation><cell mediated therapies><cell-based therapeutic><cell-based therapy><cellular therapeutic><cellular therapy><commercial application><commercial scale manufacturing><commercialization><corona virus disease 2019><coronavirus disease 2019><coronavirus disease-19><coronavirus infectious disease-19><cost><data interpretation><detection assay><developmental><diagnostic kit><experiment><experimental research><experimental study><experiments><fighting><fluid><full scale manufacturing><gene product><genome editing><genomic editing><high temperature><improved><innovate><innovation><innovative><instrumentation><interest><lab equipment><laboratory equipment><laboratory facility><laboratory technology><large scale manufacturing><large scale production><lateral flow assay><lateral flow test><liquid><mRNA><mRNA vaccine><mRNA-based vaccine><manufacture><manufacturing facility><manufacturing plants><manufacturing process><manufacturing ramp-up><manufacturing scale-up><mass production><nano particle><nano-sized particle><nanoparticle><nanosized particle><new diagnostics><new vaccines><next generation diagnostics><next generation vaccines><novel><novel diagnostics><novel vaccines><nuclease><oligos><ontogeny><particle><pharmaceutical><portability><preservation><produce vaccines><production plants><prototype><rapid detection><real world application><regulatory gene><resistant><salt><scale up batch><scale up production><social role><success><supply chain><test kit><therapeutic RNA><therapeutic vaccination><tool><trans acting element><transcriptomics><upscale manufacturing><vaccine for immunotherapy><vaccine immunotherapy><vaccine-based immunotherapy>