Factors to be considered for improving the HDR (IDT webinar)

 Factors to be considered for improving the HDR

  • Improved designing

    • Selection of high activity guides

    • Using the other type of Cas – > changing from Cas9 to Cas12a

    • Blocking mutations to prevent re-cutting after HDR

      • Introducing non-complement base in gRNA to reduce the activity of Cas9

    • Optimize homology arm length

      • Same length but different gene – > give different HDR efficiency

      • In general sense, larger – > better HDR but it will reach plateau, thus we use the lowest length which gives the best HDR (~200 bp)

  • Improved reagent

    • Small molecule to modulate pw

      • Available by the company (blind solution)

    • Modification of repair template to increase stability


Web-based tool to design the gRNA

www.idtdna.com/HDRdesigntool


Info:

www.idtdna.com/HDR

www.idtdna.com/CRISPR


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