GeneArt™ Genomic Cleavage Detection Kit

TALs or TALENs are transcription activator-like effector nuclease proteins that are naturally occurring transcriptional activators secreted by Xanthomonas spp. into their plant hosts. GeneArt TALs are derived from Xathomonas TAL effectors, the DNA-binding domain of which consists of a variable number of amino acid repeats. Each repeat contains 33–35 amino acids and recognizes a single DNA base pair. The DNA recognition occurs via 2 hypervariable amino acid residues at positions 12 and 13 within each repeat, called repeat-variable di-residues (RVDs). TAL effector repeats can be assembled in modular fashion, varying the RVDs to create a TAL protein that recognizes a specific target DNA sequence.

CRISPR-STOP is a method of inserting STOP codon sequences to generate knockouts.

Please refer to the following article: CRISPR-STOP: gene silencing through base-editing-induced nonsense mutations.

Find additional tips, troubleshooting help, and resources within our Genome Editing Support Center.

We do offer a multiple cloning site sequence in the place of the effector domain sequence for our TAL MCS entry vector. This option allows you to insert any protein-coding sequence, and allows your resulting TAL protein to deliver the effector in a sequence-specific manner anywhere in the genome. We also provide gene synthesis services to generate any effector domain for which you don't have a template.

While our Invitrogen GeneArt Precision TALs required a T at the 5´end and 13-18 bp spacing between the forward and reverse TAL effectors for proper pairing of Fok1 nucleases, the Invitrogen GeneArt PerfectMatch TALs allow for targeting of any sequence across the genome and eliminates the 5´ T constraints. Additionally, the spacing between the two effectors is optimal at 15-16 bp.

The 19 bp binding domains perform better for the nucleases. The binding sites do not need to be the same size; however, best performance for the nucleases is with the 19 bp binding domains.