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Related Concept Videos

CRISPR01:59

CRISPR

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Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
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Integral membrane proteins are tightly associated with the cell membrane and play a crucial role in cell communication, signaling, adhesion, and transport of the molecules. Some integral membrane proteins are present only in the membrane monolayer. For example, the enzyme fatty acid amide hydrolase is present in the cytoplasmic side of the membrane monolayer. In contrast, another type of integral membrane protein, also known as a transmembrane protein, spans across the membrane. Transmembrane...
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Dimensional Analysis03:40

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Dimensional analysis, also known as the factor label method, is a versatile approach for mathematical operations. The main principle behind this approach is: the units of quantities must be subjected to the same mathematical operations as their associated numbers. This method can be applied to computations ranging from simple unit conversions to more complex and multi-step calculations involving several different quantities and their units.
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For successful DNA replication, the unwinding of double-stranded DNA must be accompanied by stabilization and protection of the separated single strands of the DNA. This crucial task is performed by single-strand DNA-binding (SSB) proteins. They bind to the DNA in a sequence-independent manner, which means that the nitrogenous bases of the DNA need not be present in a specific order for binding of SSB proteins to it. The binding of SSB proteins straightens single-stranded DNA (ssDNA) and makes...
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BEST: Barcode Enabled Sequencing of Tetrads
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Protein Barcodes Enable High-Dimensional Single-Cell CRISPR Screens.

Aleksandra Wroblewska1, Maxime Dhainaut1, Benjamin Ben-Zvi2

  • 1Precision Immunology Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.

Cell
|October 23, 2018
PubMed
Summary
This summary is machine-generated.

Researchers developed a novel protein barcoding system (Pro-Codes) to overcome limitations in CRISPR gene function screening. This technology allows for high-dimensional, single-cell phenotyping, revealing new insights into cancer cell immune editing and gene regulation.

Keywords:
CRISPRT cellscancerfunctional genomicsinterferon gamma pathwaymass cytometrypooled screenprotein barcodessingle cell analysistumor immunology

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Area of Science:

  • Molecular Biology
  • Immunology
  • Biotechnology

Background:

  • CRISPR screens are vital for gene function discovery but limited by DNA barcoding resolution.
  • Current DNA-based methods restrict phenotyping capabilities and lack single-cell resolution.

Purpose of the Study:

  • To develop a protein-level barcoding system (Pro-Codes) for enhanced CRISPR screening.
  • To enable high-dimensional, single-cell phenotyping for broader functional genomics applications.

Main Methods:

  • Synthesized protein barcodes (Pro-Codes) using triplet combinations of linear epitopes.
  • Introduced Pro-Code vectors into cells and analyzed using CyTOF mass cytometry with 14 antibodies.
  • Paired Pro-Codes with CRISPR for simultaneous analysis of phenotypic markers, including phospho-signaling, in knockout cells.

Main Results:

  • Generated >100 unique Pro-Codes and detected 364 populations, creating the largest set of protein reporters.
  • Identified Psmb8 and Rtp4 as crucial for antigen-dependent immune editing in cancer cells.
  • Discovered Socs1 as a negative regulator of Programmed Death-Ligand 1 (Pd-l1).

Conclusions:

  • Pro-Code technology enables unprecedented simultaneous, high-dimensional protein-level phenotyping of hundreds of genes at single-cell resolution.
  • This advancement significantly expands capabilities for functional genomics and drug discovery research.