Non-targetingsgRNAs

[Meghana Kulkarni]

Hi,
I have several questions regarding the use and design of control non-targeting sgRNAs for any given species. Is there a tool that one can use to do this?

Also-
1. Does the GeCKO library have non-targeting sgRNAs
2. How many? What were the sequences used?
3. Any recommendations on how many non-targeting sgRNAs one should include in a library of a particular size?

Best,
Meghana

[Neville Sanjana]

Hi Meghana,

The current GeCKO library does not but we are working hard to create a new version that will have control (non-targeting) sgRNAs. I don't think there are any hard and fast rules for how many controls to have but I think you'd want a large enough number so that you have control sgRNAs that span the same range of representation as the non-controls in the library. 

hope that helps!

- Neville

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[Meghana Kulkarni]

Thanks Neville. Is there an easy way for a lay person like me to do this for human and mouse genomes i.e is there a web design tool available? This is probably not easy to do and requires considerable computational resources.

Meghana

[Neville Sanjana]

Hi Meghana,

I am happy to share our control sgRNA for human and mouse genomes. These will be publicly available soon as part of new libraries but if you'd like to have access to the control sgRNA sequences ahead of time, please email me directly and I can share them.

best,

- Neville

[Charles Lee]

Hi Neville,

Could you please seed the control sgRNA sequences to me? My Email lichang...@gmail.com. Thanks in advance!

在 2014年3月18日星期二UTC+8上午10时04分33秒，Neville Sanjana写道：

[Harmony Ma]

Hi Neville,

Could you please send the non-targeting sgRNA to me, thank you very much. 

my email harmon...@gamil.com

harmony

在 2014年3月17日星期一UTC-7下午7时04分33秒，Neville Sanjana写道：

[carla....@hospitalitaliano.org.ar]

Hi Neville

I know that someone have already asked you for those control sgRNA, I am in the same situation. So could you please send the non-targeting sgRNA to me too? gimene...@hotmail.com

Thanks a lot!!

 

[gaf...@gmail.com]

Hi Neville,

Could you please also send me your non-targeting gRNA controls for mouse.  Thank you so much!  cjga...@ucalgary.ca

Chris

[sisi2...@gmail.com]

Hi Neville:

Could you please send me the sequences too?

It's so nice for you to do that!

Si

[Andy Ng]

Dear Neville, can you send me some control (non-targeting) sgRNA.

Many thanks,

Andy

[Leigh Brody]

Hi Neville, I'm also facing the same question here. Could I please also have the control sgRNA sequences for human and mouse? My email address is leigh...@gmail.com. Thanks!

[Neville Sanjana]

Dear Leigh,

The GeCKO v2 libraries contain 1000 non-targeting (control) sgRNA sequences for human and 1000 non-targeting (control) sgRNA sequences for mouse. You can find the list of sequences as CSV files here: http://www.addgene.org/crispr/libraries/geckov2/  (look under the Add to Cart buttons for the download links)

If you just need a few controls, here are 10 for each species (a subset of the 1000 referenced above).

Good luck!

- Neville

human nontargeting guides
target_id	spacer_seq
rg_0111	ACGGAGGCTAAGCGTCGCAA
rg_0135	CGCTTCCGCGGCCCGTTCAA
rg_0143	ATCGTTTCCGCTTAACGGCG
rg_0166	GTAGGCGCGCCGCTCTCTAC
rg_0194	CCATATCGGGGCGAGACATG
rg_0240	TACTAACGCCGCTCCTACAG
rg_0241	TGAGGATCATGTCGAGCGCC
rg_0311	GGGCCCGCATAGGATATCGC
rg_0313	TAGACAACCGCGGAGAATGC
rg_0345	ACGGGCGGCTATCGCTGACT
mouse nontargeting guides
target_id	seq
rand_seq5543	GCGAGGTATTCGGCTCCGCG
rand_seq9270	GCTTTCACGGAGGTTCGACG
rand_seq4841	ATGTTGCAGTTCGGCTCGAT
rand_seq6647	ACGTGTAAGGCGAACGCCTT
rand_seq8828	GACTCCGGGTACTAAATGTC
rand_seq5431	CCGCGCCGTTAGGGAACGAG
rand_seq4112	ATTGTTCGACCGTCTACGGG
rand_seq2389	ACCCATCGGGTGCGATATGG
rand_seq8562	CGGGCGTCACCTGCTAGTAA
rand_seq1698	GCTTCTACTCGCAACGTATT