Purple Ghost
posted over 3 years ago

Custom-made CRISPR pooled gRNA KO library_Lentiviral vectors

I want to use a CRISPR library to knockout a certain number of human genes that I'm interested in.

Since I'm new in the field of genome editing, I would like to find a reliable company to generate for me the library in the form of ready-to-transduce lentiviral-vectors.

I am going to follow the Nature protocols 2017 of Joung et al. that I found really helpful, so I would like the library to be made either in lentiCRISPRv2 or lentiGuide-puro vector backbone.

My questions are:

  1. Can any of you recommend or had experience with a company that can provide custom-made libraries in a customer-provider vector backbone in the form of lentiviral vectors?
  2. In order for the Nature protocols 2017 of Joung et al., to be applied in a custom-made library what factors should I have in mind before ordering the library?
Turquoise Ghoulover 3 years ago

Please check Dharmacon. They have whole range of custom-made library.

Fuchsia Kelpieover 3 years ago

Thank you for your interest in our work. To answer your questions:

1. I do not have experience with this.

2. As long as the sgRNA target sequences use the U6 promoter and sgRNA scaffold sequences found in the lentiCRISPRv2 or lentiGuide-puro vectors, you can apply the protocols in Joung Nat Protocols 2017. For instance, you will be able to switch out the puromycin resistance gene for GFP, and still be able to apply the protocol.

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