このコンテンツを視聴するには、JoVE 購読が必要です。 サインイン又は無料トライアルを申し込む。
HOX Loci Focused CRISPR/sgRNA Library Screening Identifying Critical CTCF Boundaries
章
- 00:04標題
- 00:49The High Titer sgRNA Library Lentivirus Generation
- 01:44Titration of Lentiviral Library in MOLM13 Leukemia Cells
- 02:59Transduction of the Pooled CRISPR-Cas9 KO Library
- 04:20Screening of the Pooled CRISPR-Cas9 KO Library with One-Step RT-qPCR
- 05:41Verification of Integrated sgRNAs Positive Clones through Genotyping and Sanger Sequence
- 06:37Detection of sgRNAs Induced Indel Mutation by Nuclease Digestion Assay
- 07:48Results: Pooled CRISPR-Cas9 KO Library Screening and Identification
- 09:19Conclusion
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.
