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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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CRISPR and crRNAs02:53

CRISPR and crRNAs

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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.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
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Bode Plots Construction01:24

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The Bode plot is an essential tool in control system analysis, mapping the frequency response of a system through a magnitude plot and a phase plot, both against a logarithmic frequency axis. To construct a Bode plot, consider the transfer function H(ω):
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Construction of Root Locus01:15

Construction of Root Locus

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The construction of a root locus involves several key steps to analyze and visualize the behavior of a system's poles with varying gain. The number of branches in the root locus equals the number of closed-loop poles and is symmetrical about the real axis.
For positive gain values, the root locus exists on the real axis to the left of an odd number of finite open-loop poles or zeros. The root locus starts at the open-loop poles and traces the paths of the closed-loop poles as the gain...
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Construction of Frequency Distribution01:15

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A frequency distribution table can be constructed using the steps given below.
First, make a table with two columns—one with the title of the data that needs to be organized, and the other column for frequency. [Draw a third column for tally marks if needed]. Then, take a look at the items given in the data set and decide if an ungrouped frequency distribution table or a grouped frequency distribution table would be more suitable. If there are large sets of different values, then it is...
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The Antiviral System of Bacteria and Archaea: CRISPR01:23

The Antiviral System of Bacteria and Archaea: CRISPR

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CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats is a adaptive immune system found in bacteria and archaea that protects against viral infections. This system enables prokaryotic cells to identify, remember, and neutralize foreign genetic elements, primarily bacteriophages, by storing fragments of the invader’s DNA as a genetic memory.The CRISPR immune response begins during an initial infection. Cas (CRISPR-associated) proteins play a central role in this...
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Related Experiment Video

Updated: Feb 10, 2026

Author Spotlight: Simplifying Genome-Wide Plasmid Library Construction Using CRISPRmass
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Author Spotlight: Simplifying Genome-Wide Plasmid Library Construction Using CRISPRmass

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Construction of CRISPR Libraries for Functional Screening.

Carsten P Carstens1, Katherine A Felts2, Sarah E Johns2

  • 1Genomics R&D, Agilent Technologies Inc., La Jolla, CA, USA. carsten.carstens@agilent.com.

Methods in Molecular Biology (Clifton, N.J.)
|May 14, 2018
PubMed
Summary

This study presents a new method for creating high-quality CRISPR/Cas9 guide RNA libraries. These improved libraries enhance gene function identification by ensuring accurate and even distribution of guide RNAs for robust screening results.

Keywords:
CRISPR-CASFunctional genomicsGene editingOligo library synthesisPlasmid library construction

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Area of Science:

  • Molecular Biology
  • Genetics
  • Bioinformatics

Background:

  • CRISPR/Cas9 technology enables targeted gene knockouts and repression for gene function identification.
  • Pooled guide RNA libraries are crucial for high-throughput screening, linking genes to phenotypes.
  • Screening data quality heavily relies on the accuracy and evenness of guide RNA distribution in libraries.

Purpose of the Study:

  • To develop an improved method for constructing complex plasmid libraries for CRISPR/Cas9 screening.
  • To achieve high representation and tight distributions of guide RNAs within these libraries.
  • To enhance the reliability and accuracy of CRISPR/Cas9-based functional genomics screens.

Main Methods:

  • Construction of plasmid libraries using pooled, designed oligomers.
  • Optimization of library synthesis to ensure high member representation.
  • Characterization of library distribution using percentile ratios.

Main Results:

  • Successfully constructed plasmid libraries with over 60,000 members.
  • Achieved a tight distribution of guide RNAs, with a 95th/5th percentile ratio below 3.5.
  • Demonstrated a method for generating high-quality, complex guide RNA libraries.

Conclusions:

  • The described method enables the creation of superior plasmid libraries for CRISPR/Cas9 screening.
  • High-quality libraries are essential for accurate gene function identification and phenotype linkage.
  • This advancement improves the robustness and reliability of functional genomics studies.