The High Titer sgRNA Library Lentivirus Generation
1:44
Titration of Lentiviral Library in MOLM13 Leukemia Cells
2:59
Transduction of the Pooled CRISPR-Cas9 KO Library
4:20
Screening of the Pooled CRISPR-Cas9 KO Library with One-Step RT-qPCR
5:41
Verification of Integrated sgRNAs Positive Clones through Genotyping and Sanger Sequence
6:37
Detection of sgRNAs Induced Indel Mutation by Nuclease Digestion Assay
7:48
Results: Pooled CRISPR-Cas9 KO Library Screening and Identification
9:19
Conclusion
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Our pooled sgRNA called the library screening, is a powerful genetic approach to identify and evaluate the biological function of the noncoding elements throughout the genome. This technique allows us to examine the effect of disrupting the bindin
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A CRISPR/sgRNA library has been applied to interrogating protein-coding genes.However, the feasibility of a sgRNA library to uncover the function of a CTCF boundary in gene regulation remains unexplored. Here, we describe a HOX loci specific sgRNA library to elucidate the function of CTCF boundaries in HOX loci.