DNA insert using CRISPR

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4 years ago

Milena Alexeyeva

3

DNA insert using CRISPR

I would like to excise a large strand of DNA and insert a new one using CRISPR. My problem is that my strand will be a little over 1kb and I am not sure if this is going to be a limiting factor. Also, how long should the homology arms be for a region of this size?

Experiment: CRISPR Human - Repression miR-21

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**I.Zawadzka** · Answer 1 · 2019-12-15T20:46:29+00:00

You should be able to insert the new region without any problems. For the homologous arms you should use 800bp arms that are homologous to the regions flanking the 1kb excised sequence. Good luck!

**Nicholas Ferguson** · Answer 2 · 2020-04-29T10:43:33+00:00

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Homology arm length depends on the species of cell you want to transform, short for prokaryotes (as little as 50 bp) and up to 500 bp for eukaryotes

Answered 4 years ago

Nicholas Ferguson

**I.Zawadzka** · Answer 3 · 2019-12-15T20:46:29+00:00

You should be able to insert the new region without any problems. For the homologous arms you should use 800bp arms that are homologous to the regions flanking the 1kb excised sequence. Good luck!