Overview
The development of novel gene-editing approaches goes beyond the treatment of diseases or their symptoms. However, analytical solutions for CRISPR/Cas9 analysis need to keep up with the demands of these new approaches.
Leverage highly sensitive techniques for characterizing large synthetic guide RNAs and in vitro transcribed RNA products, and understand low-abundance post-translational modifications (PTMs) of Cas9 proteins and CRISPR/Cas9 effects on the proteome. With dedicated analytical solutions, personalized medicine is only a step away.
Workflow
Size and integrity of sgRNA and mRNA
The latest gene-editing approaches require overcoming new analytical challenges. Confirmation of the size, integrity and purity of large synthetic guiding RNAs (sgRNA and pegRNA) and confident analysis of Cas9 mRNA require the highest data quality over a wide size range.
Move past analytical challenges with high separation quality, sensitivity and superior reproducibility. Reclaim your time with intuitive software and kit-based workflows that are compatible with compliant data systems.
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Simultaneous, high-resolution measurement of sgRNA and Cas9 mRNA over a wide size range
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High repeatability and minimized secondary structure formation with stable temperature control of the capillary and cartridge
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High-sensitivity detection of analytes and low-abundance impurities
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Easy assay transfer to regulated environments due to compatibility with the Empower Chromatography Data System (CDS)
Size and integrity of sgRNA and mRNA
Solution
- High-quality resolution and sizing of RNAs and impurities
- Medium to large sample batches
BioPhase 8800 system
RNA 9000 Purity & Integrity kit
BioPhase BFS capillary cartridge
Featured resources
Learn about an intuitive kit-based solution for the characterization of mRNA integrity and purity. Break through boundaries with workflows that provide the highest resolution for sgRNA and mRNA in a single analysis.
Set your own schedule for mRNA analyses by streamlining your CE data management with the Empower CDS.
Size and integrity of sgRNA and mRNA
Solution
- High-quality resolution and sizing of RNAs and impurities
- Smaller sample sets
PA 800 Plus system
RNA 9000 Purity & Integrity kit
BFS capillary cartridge for the PA 800 Plus system
Workflow
Cas9 protein characterization with peptide mapping
A deeper understanding of gene-editing mechanisms paves the way for higher target specificity and minimized side effects. Engineering Cas9 proteins with improved binding and stability can be one approach. However, ensuring the correct function requires comprehensive characterization of the Cas9 proteins.
Obtain high-quality, comprehensive sequence and fragment coverage with advanced MS/MS sensitivity. Explore how to elucidate the position and identity of amino acid substitutions/isomers and other challenging PTMs with confidence.
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High sequence coverage and comprehensive PTM determination with high-quality MS/MS collision-induced dissociation (CID)
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Identification of low-abundance amino acid mutations using the Zeno trap
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Determination of identity and position of amino acid isomers and other challenging PTMs with electron activated dissociation (EAD)
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High fragment coverage for long, difficult-to-fragment peptides using Zeno EAD
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Short learning curve with intuitive SCIEX OS software for acquisition
Cas9 protein characterization with peptide mapping
Solution
- Differentiation of amino acid isomers and identification of challenging PTMs
- Most sensitive detection of low abundance PTMs and impurities
- Meeting flexibility needs to perform a range of additional workflows
ZenoTOF 7600 system
Biologics Explorer software
SCIEX OS software
Featured resource
Unleash the potential of protein characterization with novel fragmentation solutions. Explore paths to understanding complex PTMs of your engineered Cas9 proteins with unprecedented depth.
Cas9 protein characterization with peptide mapping
Solution
- Straightforward confirmation of sequence and identification of PTMs
- Robust, analytical flow setup
ExionLC AE system
X500B QTOF system
Biologics Explorer software
SCIEX OS software
Workflow
pegRNA analysis with CGE
More sophisticated, chemically synthesized gRNAs are in development with the goal of increasing efficiency and specificity of CRISPR/Cas9 gene editing. One of these newly emerging classes is pegRNA with sizes ranging from around 120–250 nucleotides (nt).
The length and associated risk of secondary structure formation, however, pose challenges for the analytical assessment of its purity.
Move beyond these boundaries of CRISPR/Cas9 analysis and confirm size and purity for large pegRNA with confidence. Rely on high resolving power and repeatability to separate your products from impurities with similar properties.
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Reliable purity assessment through high-resolution separation of full-length pegRNAs from their impurities
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Control of secondary structures of chemically synthesized pegRNAs (>100 nt)
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Excellent repeatability with accurate and stable temperature control
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Easy assay transfer to regulated environments due to compatibility with the Empower CDS
pegRNA analysis with CGE
Solution
PA 800 Plus system
ssDNA 100-R kit
Workflow
sgRNA intact mass analysis
Oligonucleotides—such as guide RNAs, sgRNAs and pegRNAs—are prone to a high number of impurities due to their stepwise chemical synthesis. Obtaining molecular weight information to confirm the identity of the main product and quantitation of related impurities is key for quality assessment prior to performing lengthy cell-based assays.
Take control of the purity of your products with high-quality, accurate mass data derived from intuitive solutions. Enable your team to gain key insights that are relevant for ensuring safety and efficacy.
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Excellent spectra quality with superior ionization and declustering of salt adducts using the Turbo V ion source
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A short learning curve with intuitive SCIEX OS software for acquisition and processing
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Additional quantitation and deconvolution options with SCIEX OS software
sgRNA intact mass analysis
Solution
- Straightforward molecular weight determination of sgRNAs and impurities
- Robust, analytical flow setup
ExionLC AE system
X500B QTOF system
SCIEX OS software
sgRNA intact mass analysis
Solution
- Straightforward molecular weight determination of sgRNAs and impurities
- Detection of low abundance impurities with superior linear dynamic range (LDR)
ExionLC AE system
ZenoTOF 7600 system
SCIEX OS software
Featured resource
Imagine taking control of your CRISPR/Cas9 gene-editing systems by ensuring sgRNA integrity and purity. Be empowered to take on the challenge of future medicine development with accurate mass solutions that provide meaningful answers.
Workflow
On/off target effects
Despite efforts to increase the specificity and efficiency of CRISPR/Cas9 gene editing, unexpected effects on the proteome can occur. Biochemistry-based assays are blind towards such changes, or they only provide limited information on affected proteins.
Give your team the advantage of comprehensive unbiased characterization of proteome changes to streamline the development of gene-editing tools. Reach new milestones with the dynamic range, sensitivity and reproducibility needed to better understand on- and off-target effects of gene therapies while enabling simultaneous quantitation in samples with limited amounts.
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A deeper understanding of CRISPR/Cas9 effects on the proteome with high MS/MS spectral quality
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Confident identification of even low-abundance species with the Zeno trap
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Identification of off-target effects for comprehensive characterization
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A short learning curve with intuitive SCIEX OS software for acquisition
On/off target effects
Solution
M5 MicroLC system
ZenoTOF 7600 system
SCIEX OS software
Featured resources
Clearly understand CRISPR/Cas9 effects on the proteome. Leverage high-quality MS/MS spectral data to identify even low-abundant, unexpected changes that ligand-binding assays might miss.
Master the development of gene-editing approaches with the ability to confidently detect proteome changes that are often missed by traditional assays. Access the highest quality spectral data to ensure the safety and efficacy of your next-generation therapies.
All resources
Clearly understand CRISPR/Cas9 effects on the proteome. Leverage high-quality MS/MS spectral data to identify even low-abundant, unexpected changes that ligand-binding assays might miss.
Master the development of gene-editing approaches with the ability to confidently detect proteome changes that are often missed by traditional assays. Access the highest quality spectral data to ensure the safety and efficacy of your next-generation therapies.
Create pathways for effective method development and learn how to achieve the highest quality data for RNA products with sizes ranging from 50–9,000 nucleotides and beyond.
Unleash the potential of protein characterization with novel fragmentation solutions. Explore paths to understanding complex PTMs of your engineered Cas9 proteins with unprecedented depth.
Overcome secondary structure challenges presented by large-sized oligonucleotides for gene editing. Learn how to enable accurate and easy purity analysis of complex synthetic pegRNAs.
Imagine taking control of your CRISPR/Cas9 gene-editing systems by ensuring sgRNA integrity and purity. Be empowered to take on the challenge of future medicine development with accurate mass solutions that provide meaningful answers.
Learn about an intuitive kit-based solution for the characterization of mRNA integrity and purity. Break through boundaries with workflows that provide the highest resolution for sgRNA and mRNA in a single analysis.
Set your own schedule for mRNA analyses by streamlining your CE data management with the Empower CDS.
Associated applications
Lipid nanoparticles and non-viral carriers
Expand the possibilities of drug delivery with innovative analytical solutions for lipid raw materials, LNPs and other non-viral carriers. Move beyond boundaries in structural elucidation and monitoring of analytes to estabilish confidence in the quality and stablity of your products.
Subunit mass analysis and middle-down
Take control of the unknowns. Gain confident sequence confirmation and localization of post-translational modifications (PTMs) at the sub-unit level.