After decades of development and innovation, Creative Biolabs is committed to demonstrating our strong expertise and capabilities in the field of ribosome research, as well as expanding our research and service capabilities to multiple fields. This five-part slide we will briefly introduce ribosome analysis technology, polysome profiling, ribosome profiling, ribosome affinity purification, and highlight the ribosome analysis solutions provided by Creative Biolabs.
5. 标题内容
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King, Helen A., and André P. Gerber. "Translatome profiling: methods for genome-scale
analysis of mRNA translation." Briefings in functional genomics 15.1 (2016): 22-31.
Experimental
approaches to
studying the
translatome
7. Polysome Profiling
标题内容
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Karamysheva, Zemfira N., et al. "Polysome profiling in Leishmania, human cells and mouse testis." JoVE (Journal of
Visualized Experiments) 134 (2018): e57600.
Lysate preparation
Sucrose gradient preparation and
ultracentrifugation
Analysis of the fractions
Polysome fractionation and sample
collection
9. 标题内容
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Bagheri, Atefeh, et al. "Tracing Translational Footprint by Ribo-Seq: Principle, Workflow, and Applications to Understand the Mechanism of
Human Diseases." Cells 11.19 (2022): 2966.
Ribosome Profiling (Ribo-Seq)
10. 标题内容
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Data analysis
Freezing ribosomes on mRNAs to avoid ribosome run-off
Ribonuclease digestion breaks down unprotected regions of mRNA
mRNA–ribosome complex recovery and RNA purification
Library generation and deep sequencing
Key Steps of Ribosome Profiling
12. 标题内容
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Heiman, Myriam, et al. "Cell type–specific mRNA purification by translating ribosome affinity purification (TRAP)." Nature protocols 9.6 (2014): 1282-1291.
(a) The cell type of interest is targeted with
appropriate genetic elements to express the
EGFP-L10a transgene.
(b) The translating polyribosomes (polysomes)
originating from nontargeted cells (gray cells) do
not have an EGFP tag on their ribosomes.
(d) Only the tagged polysomes are captured on an
anti-GFP affinity matrix, which can be used for
purification of the cell type-specific mRNA
associated with tagged polysomes.
(c) Those originating from targeted cells (green
cells) have an EGFP tag on their ribosomes.
Ribosome Affinity Purification (RAP, or Translating RAP (TRAP))
Strategy
13. 标题内容
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标题内容
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Method Advantages Disadvantages Optimal Applications
Polysome Profiling
• Ribosome density measurement
• Fractionation by ribosome occupancy
on mRNA
• Specialized equipment
• Larger number of cells (∼107 )
• Labor intensive
• Pseudopolysome contamination
• Lack of cell-type specificity
• Cultured cells/tissues
• Initial screen for translational changes
• To track the translational status of a given
mRNA/particular set of mRNA with known
identity
Ribosome Profiling
• Ribosome density measurement
• Ribosome positional information
• The discovery of new uORFs,
alternative start codons, alternative
protein isoforms, decoding speeds,
translational pause sites, and
translational dynamics
• Specialized equipment
• Larger number of cells (∼107 )
• Labor intensive
• PseudoRPF contamination
• Expensive (deeper sequencing
required due to fragment size)
• Extensive bioinformatics—annotation
of small sequencing fragments
• Lack of cell-type specificity
• Cultured cells/tissues
• Dissection of initiation and elongation
events
• Determination of molecular mechanisms
of translational control
Ribosome Affinity Purification
• Smaller number of cells (< 106 )
• Cell/tissue-type specific
• Simple, cheap, and robust
• Genetic modification, e.g., transgenes
to express tagged RPs
• Lower resolution
• Samples with genetically tagged
ribosomes
Advantages, Disadvantages, and Optimal Applications for the Three
Translatome Analysis Techniques
14. Ribosome
Analysis at
Creative Biolabs
PART FIVE
Ribosome Analysis Technology Polysome Profiling Ribosome Profiling Ribosome Affinity Purification Ribosome Analysis at Creative Biolabs
15. Creative Biolabs is a world-leading CRO and recognized expert dedicated to providing high-quality services to global customers.
Benefitting from the development and accumulation, Creative Biolabs is equipped with cutting-edge instruments, advanced technology
platforms, as well as a professional technical team. Creative Biolabs is committed to demonstrating our strong expertise and capabilities
in the field of ribosome research and expanding our research and service capabilities to ther areas.
What Can We Do
Ribosomal Marker Antibody
Development Services
Ribosome Separation and
Extraction Services
Ribosome Analysis
Services
17. Ribosomal Transcriptome Ribosomal Proteomics
The conventional belief that a ribosome is merely a
translation machine has significantly impeded
advances in ribosomal proteomics. With the proposal
of the ribosome filtering hypothesis and the study of
ribosome-specific regulation ability, ribosomal
proteomics has become a powerful tool for ribosome
research.
Microarray Technology
RNA-Seq
Both biological sciences and medicine rely on rapid
genomic and transcriptomic analysis techniques to
probe cellular state, physiology, and activity. The
development of ribosomal transcriptomics has laid
the foundation for further understanding of the
protein synthesis process.
Label-Dependent Strategies
Label-Independent Strategies
Ribosome Analysis Services
18. 标题内容
请在此输入详细内容
Lessard, Frédéric, Léa Brakier‐Gingras, and Gerardo Ferbeyre. "Ribosomal proteins control tumor suppressor pathways in response to nucleolar stress."
Bioessays 41.3 (2019): 1800183.
Ribosomal Marker Antibody Development Services
19. Why Choose Us
Perfect experimental
platform
We have established a
complete experimental
platform for ribosome
purification and analysis.
Professional
research team
Our excellent research
team specializes in
theoretical knowledge
and practical skills in
ribosome isolation and
analysis.
Abundant
experience
We have extensive
experience from
numerous successful
ribosome projects. We
can tailor a professional
process to your project
requirements.
One-stop service of
high quality
Our team can provide
diverse and low-cost,
high-efficiency one-stop
ribosome analysis
services.
