
ssODN-mediated knock-in with CRISPR-Cas for large genomic …
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).
ssDNA和ssODNs - 知乎 - 知乎专栏
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。
ssODN Strategies for Large Fragment Insertion via CRISPR
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. ...
ssODN-mediated knock-in with CRISPR-Cas for large genomic ... - PubMed
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.
Highly efficient genome editing for single-base substitutions using ...
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,...
Mechanistic and genetic basis of single-strand templated ... - Nature
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...
Efficient ssODN-Mediated Targeting by Avoiding Cellular …
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 …
Efficient In Vivo Introduction of Point Mutations Using ssODN and …
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.
Efficient ssODN-Mediated Targeting by Avoiding Cellular ... - PubMed
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.
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