2016年1月20日 · Here we show efficient gene KI in rats by combining CRISPR-Cas with single-stranded oligodeoxynucleotides (ssODNs). First, a 1-kb ssODN co-injected with guide RNA (gRNA) and Cas9 messenger RNA...

The ssODN is comprised of three parts: two homology arms sandwiching the region to be edited. The 5' homology arm (HA): this homology arm will encompass the ~50 nt upstream of the Cas9 cleavage site. The Cas9 cleavage site is 3 nt upstream of the PAM site (see diagram below).

2023年7月6日 · ssODN ：寡核苷酸（single-stranded oligonucleotides） ssODNs包含一个24nt长的非同源中心区域， 以取代guide RNA ( gRNA ) 靶向的基因组DNA序列 ，同时在FKB12的3个阅读框中引入终止密码子，其两侧有短的 (<75 nt)同源序列。

2025年3月10日 · At the core of ssODN-mediated gene editing is homology-directed repair (HDR), a natural process that ssODNs exploit to introduce genetic changes. When a double-strand break (DSB) is induced in the DNA, typically by CRISPR-Cas9, the cell’s repair machinery can use an ssODN as a template to repair the break, incorporating the desired sequence. ...

2016年1月20日 · Here we show efficient gene KI in rats by combining CRISPR-Cas with single-stranded oligodeoxynucleotides (ssODNs). First, a 1-kb ssODN co-injected with guide RNA (gRNA) and Cas9 messenger RNA produce GFP-KI at the rat Thy1 locus.

2019年3月18日 · To improve the efficiency of knock-in with single-stranded oligo DNA nucleotides (ssODNs), we have investigated optimal design of ssODNs in terms of the blocking mutation, orientation, size,...

2021年11月19日 · Single-stranded oligodeoxynucleotides (ssODNs) are widely used as DNA repair templates in CRISPR/Cas precision genome editing. However, the underlying mechanisms of single-strand templated...

Combined with CRISPR-Cas9 technology and single-stranded oligodeoxynucleotides (ssODNs), specific single-nucleotide alterations can be introduced into a targeted genomic locus in induced pluripotent stem cells (iPSCs); however, ssODN knockin frequency is …

2020年7月14日 · We successfully applied the CRISPR-based white co-conversion strategy with a ssODN template, instead of the originally described dsDNA plasmid, to create genetically modified Drosophila melanogaster strains. We used the technique to easily introduce point mutations in two distinct chromosomes.

2021年4月13日 · Combined with CRISPR-Cas9 technology and single-stranded oligodeoxynucleotides (ssODNs), specific single-nucleotide alterations can be introduced into a targeted genomic locus in induced pluripotent stem cells (iPSCs); however, ssODN knockin frequency is low compared with deletion induction.