CRISPR/Cas9 optimized workflow

 

CRISPR Cas9

 

 

 

 

e.g. biomers.net >


Liquid Media >

Antibiotics >

 

 

Taq DNA Polymerase S >

dNTPs >

Total RNA Purification Kit >

 

 

Cas9 Protein >>

Transfection reagent GenaxxoFect >

Optimized transfection protocol for CRISPR/Cas9 methods >

 

 

Standard Agarose LE

<RealTime/qPCR Master mixes >

 

 

 

 

 

Scheme according to: Xiquan Liang et al, Journal of Biotechnology, Vol. 208, 44-53, 2015. >

 

Referenzen:

Lander ES (Jan 2016). "The Heroes of CRISPR". Cell 164 (1-2): 18–28. doi:10.1016/j.cell.2015.12.041 >. PMID 26771483.

Mojica FJ, Díez-Villaseñor C, Soria E, Juez G (Apr 2000). "Biological significance of a family of regularly spaced repeats in the genomes of Archaea, Bacteria and mitochondria". Molecular Microbiology 36 (1): 244–6. doi:10.1046/j.1365-2958.2000.01838.x >. PMID 10760181.open access publication.

Zuris JA, Thompson DB, Shu Y, Guilinger JP, Bessen JL, Hu JH, Maeder ML, Joung JK, Chen ZY, Liu DR. Cationic lipid-mediated delivery of proteins enables efficient protein-based genome editing in vitro and in vivo. Nat Biotechnol. 2015 Jan;33(1):73-80. doi: 10.1038/nbt.3081. Epub 2014 Oct 30 >.

Sojung Kim, Daesik Kim, Seung Woo Cho, Jungeun Kim and Jin-Soo Kim. Highly efficient RNA-guided genome editing in human cells via delivery of purified Cas9 ribonucleoproteins. Published in Advance April 2, 2014, doi: 10.1101/gr.171322.113. Genome Res. 2014. 24: 1012-1019 >.

Kent G. Golic. RNA-Guided Nucleases: A New Era for Engineering the Genomes of Model and Nonmodel Organisms. Genetics October 2, 2013 vol. 195 no. 2 303-308; DOI: 10.1534/genetics.113.155093 >

Liang X, Potter J, Kumar S, Zou Y, Quintanilla R, Sridharan M, Carte J, Chen W, Roark N, Ranganathan S, Ravinder N, Chesnut JD. Rapid and highly efficient mammalian cell engineering via Cas9 protein transfection. J Biotechnol. 2015 Aug 20;208:44-53. doi: 10.1016/j.jbiotec.2015.04.024. Epub 2015 May 21. >

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