sgRNA design for CRISPR-Cas9
Hi William Guillermo Vilchez Cruz
I would recommend you the attached Genscript handbook
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See the attached file for an illustration of this problem. Consider an elastic sheet S (orange) that’s laying atop a solid base B (blue). The sheet is under tension (arrows) and is held to the...
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28 May 2024 3,097 2 View
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I am just trying to find out if Anser Press is legit-there is a journal "Economic Analysis Letters" that wants me to referee for them.
18 May 2024 5,886 2 View
I am trying to clone my target sequence for CRISPR/Cas9 work. I have performed digestion and gel purificatoon of the lentiCRISPRv2 plasmid with a good yield and purity. However, when I performed...
26 March 2024 6,376 2 View
We use synthetic gRNA from IDT but we think have some problems for sgRNA hybridization with Cas9. any suggestion is welcomed!
25 March 2024 1,536 3 View
I'm doing CRISPRi with dead spCas9 on mouse cells, and normally we do the following for gRNA design: 1. 20 nt spacer 2. Extra G on the 5' to enhance expression from hU6 promoter. Final product:...
25 February 2024 4,498 3 View
Hi Everyone! I am currently designing primers to validate my CRISPR-KO mammalian cell line using TIDE. The primer is designed about 250bp upstream of the PAM site. Then, Sanger sequencing is...
27 January 2024 2,219 1 View
I am interested in knowing if there is any sgRNA library designed against MDCK cell line which is a canine cell line. i believe it shares about 84% homology with human cell line. let me know if...
03 January 2024 5,148 0 View
Lentiviruses are often used as delivery vectors for in vivo crispr screening because the lentiviral genome is able to integrate into the cellular genome, so is it okay to use AAV, a virus that...
11 December 2023 7,742 0 View
I work in a project where we need to knockout a gene at primary T cells targeting an exon that is shared between all variants. However, while the sgRNA design has been optimised ,we still see low...
24 October 2023 5,722 1 View
hi i'm gonna use crispra system for my study, how can i validate sgRNA not having off-target effect?
13 October 2023 6,733 0 View
I am trying to transfect HEK293 cells with dCas9-KRAB-DNMT3A carrying plasmid and multiple IVT sgRNAs. RNAi max invitrogen and Hiperfect Qiagen are only good for small RNAs but not plasmids. Does...
04 October 2023 3,360 1 View
Hello! I am trying to design sgRNAs for dCas9 fluorescent tagging, but all of the tools I find online are for standard CRISPR knocking in/out. Anyone have a recommended tool for dCas9?
27 August 2023 1,966 0 View
