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Related Concept Videos

Ribosome Profiling02:24

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Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
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Rapid Molecular Profiling of Defined Cell Types Using Viral TRAP.

Alexander R Nectow1, Maria V Moya2, Mats I Ekstrand3

  • 1Laboratory of Molecular Genetics, Howard Hughes Medical Institute, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA; Princeton Neuroscience Institute, Princeton University, Lot 20 Washington Road, Princeton, NJ 08544, USA.

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|April 20, 2017
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Summary

Researchers developed a viral TRAP (vTRAP) method using adeno-associated virus (AAV) to profile specific cell types in the brain. This technique allows for the detailed molecular analysis of Cre-recombinase expressing cells in various tissues.

Keywords:
AAVMCHNtsr1RNA-seqTRAPcorticothalamicmolecular profiling

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Area of Science:

  • Neuroscience
  • Molecular Biology
  • Genetics

Background:

  • Cell type characterization in complex tissues like the brain is challenging.
  • Translational profiling offers a way to study cell-specific gene expression.

Purpose of the Study:

  • To develop a versatile strategy for spatiotemporally restricted cell type profiling in the central nervous system (CNS).
  • To enable molecular dissection of specific cell populations using translating ribosome affinity purification (TRAP).

Main Methods:

  • Engineered a Cre-dependent adeno-associated virus (AAV) expressing an EGFP-tagged ribosomal protein (AAV-FLEX-EGFPL10a).
  • Utilized translating ribosome affinity purification (TRAP) to capture translating mRNAs.
  • Applied viral TRAP (vTRAP) to genetically and anatomically defined neural populations expressing Cre recombinase.

Main Results:

  • Demonstrated vTRAP's ability to target various Cre-expressing neural populations.
  • Recapitulated molecular profiles obtained by bacTRAP in corticothalamic neurons.
  • Showcased the elucidation of regional gene expression differences within cell types via spatially restricted AAV injections.

Conclusions:

  • Established the broad applicability of the vTRAP strategy for molecular dissection of CNS and peripheral cell types.
  • vTRAP provides a powerful tool for understanding cell-specific gene expression in complex biological systems.