(2014). LentiCRISPRv2 and lentiGuide-Puro: lentiviral CRISPR/Cas9 and single guide RNA CRISPR Genome-scale CRISPR-Cas9 knockout screening in human cells. Science (New York, N.Y.), 343, 84–7. Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/24336571 Reade...
内容提示: LentiCRISPRv2 and lentiGuide-Puro: lentiviral CRISPR/Cas9 and single guide RNA ? CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) is a microbial nuclease system involved in defense against invading phages and plasmids. CRISPR loci in microbial hosts contain a combination of ...
virus. lentiGuide-Puro produces >100X higher titer virus over lentiCRISPRv1 and should be used in cell lines where Cas9 has already been integrated in (e.g. using the separate lentiCas9-Blast lentivirus). For applications where Cas9 cannot first be introduced (e.g. primary cells), lenti...
lentiGuide-Puro (two vector system): 这个质粒只有SgRNA,没有Cas9元件,必须配合 lentiCas9-Blast (a separate lentiviral construct that delivers hSpCas9 and blasticidin resistance) 使用。该质粒使用方法就是预先在你的靶细胞系中转染lentiCas9-Blast,通过单克隆培养,使你的细胞系表达Cas9蛋白,然后再转染lentiGuide-...
CRISPR/Cas9 Gene Editing in Mammalian Cells Using LentiCRISPRv2/LentiGuide-Puro Vectors Different protocols have been provided; however, the success of CRISPR/Cas9 experiments is challenging. Here, we aim to describe the use of lentiviral vectors (lentiCRISPRv2/lentiGuide-Puro) for CRISPR/Cas9 ...
Here, we aim to describe the use of lentiviral vectors (lentiCRISPRv2/lentiGuide-Puro) for CRISPR/Cas9 genome editing and to provide strategies for minimizing off-targets. Troubleshooting advice will be provided based on experimental evidence. These guidelines will enable researchers and those with ...