Project description:We recently developed an easy, efficient and scalable method for tagging and live cell imaging of non-repetitive, endogenous chromosome regions via CRISPR/Cas9 mediated knock-in of a TetO repeat. For this purpose, we created optimized and irregular 48-mer and 96-mer TetO repeats. Since it is known that repetitive regions in the human genome can induce H3K9me3-mediated heterochromatin formation, we tested whether 48-mer and/or 96-mer TetO repeats induce H3K9me3 flanking their insertion sites. Using a newly developed method called as CUT&RUN, we showed that there was no significant difference in the H3K9me3 pattern flanking the insertion sites of TetO repeats when compared to wild-type cells.
2018-05-22 | GSE113832 | GEO
Project description:A universal system for streamlined genome integrations with CRISPR-associated transposases
Project description:Tagmentation is an efficient approach for sequencing library preparation, which combines DNA fragmentation by transposition of sequencing adapters, using the bacterial cut-and-paste transposase Tn5. Here we present an open-source protocol for the generation of multi-purpose hyperactive Tn5 transposase, including its benchmarking in CUT&Tag, bulk and single-cell ATAC-seq. The OpenTn5 protocol yields multi-milligram quantities of pG-Tn5E54K, L372P protein per liter of E. coli culture, sufficient for thousands of tagmentation reactions and the enzyme retains activity in storage for more than a year.
Project description:Tagmentation is an efficient approach for sequencing library preparation, which combines DNA fragmentation by transposition of sequencing adapters, using the bacterial cut-and-paste transposase Tn5. Here we present an open-source protocol for the generation of multi-purpose hyperactive Tn5 transposase, including its benchmarking in CUT&Tag, bulk and single-cell ATAC-seq. The OpenTn5 protocol yields multi-milligram quantities of pG-Tn5E54K, L372P protein per liter of E. coli culture, sufficient for thousands of tagmentation reactions and the enzyme retains activity in storage for more than a year.
Project description:Tagmentation is an efficient approach for sequencing library preparation, which combines DNA fragmentation by transposition of sequencing adapters, using the bacterial cut-and-paste transposase Tn5. Here we present an open-source protocol for the generation of multi-purpose hyperactive Tn5 transposase, including its benchmarking in CUT&Tag, bulk and single-cell ATAC-seq. The OpenTn5 protocol yields multi-milligram quantities of pG-Tn5E54K, L372P protein per liter of E. coli culture, sufficient for thousands of tagmentation reactions and the enzyme retains activity in storage for more than a year.
2025-09-02 | GSE273908 | GEO
Project description:Precise and efficient CD47 editing using hypercompact CRISPR/Cas12f