MDA-MB-231 cells were transfected with CRISPR/Cas9 and HDR plasmids targeting SNAI1 (Santa Cruz Biotechnology Inc., CA, USA). A pool of three plasmids, each encoding one guide RNA targeting exon-1, exon-2 or exon-2/intron-2 junction, was co-transfected with the Cas9 nuclease and HDR plasmids. Two days post‐transfection, cells were selected with 4 μg/ml puromycin (Merck/Millipore, Stockholm, Sweden) and single-cell colonies were expanded. Knockout clones were validated using immunoblotting and quantitative RT-PCR to analyze the mRNA levels of each of the three SNAI1 exons.
Crispr cas9 and hdr plasmids
Manufactured by Santa Cruz Biotechnology
Sourced in United States
CRISPR/Cas9 and HDR plasmids are genetic engineering tools used for gene editing. The CRISPR/Cas9 system enables targeted modifications of DNA sequences, while HDR plasmids facilitate homology-directed repair of DNA breaks. These products provide researchers with the ability to precisely manipulate genetic material in various applications.
Automatically generated - may contain errors
3 protocols using crispr cas9 and hdr plasmids
1
CRISPR/Cas9-Mediated SNAI1 Knockout
2
CRISPR/Cas9 Knockout of HMGCR Gene
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were transformed with CRISPR/Cas9 and HDR plasmids targeting HMGCR (Santa Cruz Biotechnology Inc., CA, USA). Cells were cotransfected with Cas9 nuclease and HDR plasmids, each encoding a guide RNA targeting exon 1, exon 2, or the exon 2/intron 2 junction. two days after transfection, cells were chosen with 4 µg/ml puromycin (Merck/Millipore, Stockholm, Sweden), and single-cell colonies were expanded. Knockout levels were verified using WB and qRT-PCR.
3
Gene Knockdown Using CRISPR/Cas9 and siRNA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Knockdown of STT3A, STT3B, TOP2A, and TOP2B was performed using CRISPR/Cas9 and HDR plasmids (Santa Cruz) according to the manufacturer’s instructions. Briefly, the cells were co-transfected with CRISPR/Cas9 and HDR plasmids. After 2 days, the cells were selected with puromycin for 1 week and subjected to analyses. A control CRISPR/Cas9 plasmid encoding a non-specific 20 nt guide RNA (Santa Cruz) was used as a negative control. β-catenin (CTNNB1) knockdown was performed by siRNA (Sigma). The target sequences are listed in Supplementary Table 2 .
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!