First, recognize shRNA: In studying gene function, RNAi has become a powerful tool in biological experiments because it can specifically silence genes or reduce expression. Among them, shRNA is a DNA molecule that can be cloned into an expression vector and expresses an siRNA duplex. Today, I will share two ways to quickly design shRNA. Second, the design of shRNA: 1. Sigma website Sigma has made a shRNA library for humans and mice, and the shRNA sequences of some genes have been validated, so it is most convenient to use Sigma's proven RNAi sequences directly. First open the URL: http:// The following interface appears: Take the NLRP3 gene as an example, directly enter the gene name, click search, the following interface appears: Find and click on the shRNA option in Products, the following interface appears: Click on the “Price†in the “MISSION shRNA Lentiviral Transduction Particles†line. A pop-up screen will display the shRNA for the target gene. The interface is as follows: Note: It is recommended to select the shRNA in the CDS region, 3UTR is basically not selected, and each shRNA selected should be BLAST as much as possible on the NCBI to ensure the specificity of the target. 2. Life Technologies website Life Technologies has a very useful online shRNA design software that is very easy to operate, and the designed shRNAs don't need to be Blast on NCBI. First open the URL: https://rnaidesigner.thermofisher.com/rnaiexpress/sort.do, select shRNA in the Target Design Options, take the NLRP3 gene as an example, enter the Accession number or Nucleotide sequence, the other conditions are unchanged: Click on "RNAi Design" and the following page appears: Selection principle: 1. Avoid selecting high GC sequences such as GGG and CCC; 2. The first base of the ShRNA target sequence must be G to ensure RNA polymerase transcription. If the selected destination sequence does not begin with G, a G must be added upstream of the tightness chain. According to the Rank star, select the appropriate target sequence (generally 4, one at each position), click Design shRNA Oligos: Then select the appropriate sequence in the "Default Loop Sequence", enter the appropriate loop loop sequence in the "Custom Loop Sequence", and click Design to get the shRNA sequence. Attachment: The dimension of the shRNA sequence is as follows: The best choice for Analog hearing aids with earbuds design TWS Hearing aids,TWS Hearing amplifier,TWS Hearing Earbuds Shenzhen Sunshine Technology Co.,Ltd , https://www.yatwinsz.com
Teach you to design shRNA