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sgRNA Pool for Streptococcus pneumoniae
(Pooled Library #170432)

  • Purpose

    This concise CRISPR pooled library contains only 1,499 sgRNAs to target the whole genome of S. pneumoniae D39V with one sgRNA per operon or gene. The sgRNA pool also covers core operons of other pneumococcal strains:

    • R6 (estimated 90.3% coverage of annotated genetic elements)
    • TIGR4 (72.2% coverage)
    • Hungary 19A-6 (66.6% coverage)
    • Taiwan 19F-14 (71.6% coverage)
    • 11A (66.8% coverage)
    • G54 (72.1% coverage)

    This library can be transformed into S. pneumoniae with inducible dCas9 to make an inducible CRISPR inhibition library.

  • Vector Backbone

    pPEPZ-sgRNAclone - does not express Cas9

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 170432 sgRNA pool for Streptococcus pneumoniae 1 $350 Add to Cart
Available to Academic and Nonprofits Only
  • A Cas9 plasmid is NOT included with this item and will have to be ordered separately. Can be used in conjunction with pJWV102-PL-dCas9 (Addgene #85588) and pPEPY-PF6-lacI (Addgene #85589) to generate an inducible CRISPR interference system.

Library Details

  • Species
    S. pneumoniae
  • Genes targeted
    2,111 genetic elements
  • gRNAs
    1,499

Library Shipping

This library is delivered as suspended DNA in a microcentrifuge tube on blue ice. The tube's contents will not necessarily be frozen. For best results, minimize freeze-thaws.

  • Volume
    ∼20µL
  • Concentration
    50ng/µL

Depositor Comments

The depositing lab developed crispery to analyze the NGS data: Crispery was developed in Python to streamline the initial sequencing data analyses in a single step. It automatically and easily performs read quality filtering and sgRNA alignment for the input fastq.gz files. The program returns a CSV file with all absolute sgRNA read counts per condition for downstream analysis.

Crispery is available as a compiled executable file requiring no Python installation, and can be run as a standalone program in MS Windows, macOS and Linux (Ubuntu). In addition, it is available as a PyPI module ( https://pypi.org/project/crispery/). All versions are available for download at Zenodo (https://zenodo.org/record/5226819). The original Python 3 code, user instructions, and test data are available on GitHub ( https://github.com/veeninglab/Crispery).

The depositing lab developed an R based pipeline for sgRNA design: https://github.com/veeninglab/CRISPRi-seq

How to cite this pooled library ( Back to top )

These pooled libraries were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    sgRNA pool for Streptococcus pneumoniae was a gift from Jan-Willem Veening (Addgene #170432)
  • For your References section:

    Exploration of Bacterial Bottlenecks and Streptococcus pneumoniae Pathogenesis by CRISPRi-Seq. Liu X, Kimmey JM, Matarazzo L, de Bakker V, Van Maele L, Sirard JC, Nizet V, Veening JW. Cell Host Microbe. 2021 Jan 13;29(1):107-120.e6. doi: 10.1016/j.chom.2020.10.001. PubMed 33120116