Improved in vivo gene knockout with high specificity using multiplexed Cas12a sgRNAs

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Author notes

1. Jun Zhou

   Present address: The Affiliated XiangTan Central Hospital of Hunan University, School of Biomedical Sciences, Hunan University, Changsha, 410082, China

2. Florian Heigwer

   Present address: Department of Life Sciences and Engineering, University of Applied Sciences Bingen, Bingen am Rhein, Germany

Authors and Affiliations

1. Division of Signaling and Functional Genomics, German Cancer Research Center (DKFZ), Heidelberg, Germany

   Fillip Port, Martha A. Buhmann, Jun Zhou, Mona Stricker, Alexander Vaughan-Brown, Ann-Christin Michalsen, Eva Roßmanith, Amélie Pöltl, Lena Großkurth, Julia Huber, Laura B. Menendez Kury, Bea Weberbauer, Maria Hübl, Elli Puscher, Florian Heigwer & Michael Boutros

2. Institute for Human Genetics, Medical Faculty Heidelberg, Heidelberg University, Heidelberg, Germany

   Fillip Port, Martha A. Buhmann, Jun Zhou, Mona Stricker, Alexander Vaughan-Brown, Ann-Christin Michalsen, Eva Roßmanith, Amélie Pöltl, Lena Großkurth, Julia Huber, Laura B. Menendez Kury, Bea Weberbauer, Maria Hübl, Elli Puscher, Florian Heigwer & Michael Boutros

3. Department of Cell and Molecular Biology & BioQuant, Heidelberg University, Heidelberg, Germany

   Fillip Port, Martha A. Buhmann, Jun Zhou, Mona Stricker, Alexander Vaughan-Brown, Ann-Christin Michalsen, Eva Roßmanith, Amélie Pöltl, Lena Großkurth, Julia Huber, Laura B. Menendez Kury, Bea Weberbauer, Maria Hübl, Elli Puscher, Florian Heigwer & Michael Boutros

4. Molecular Biosciences/Cancer Biology Program, Heidelberg University and German Cancer Research Center (DKFZ), Heidelberg, Germany

   Martha A. Buhmann

Contributions

F.P. and M.B. conceived the study. F.P., M.A.B., and J.Z. designed, performed and analyzed experiments. M.S., A.V.B., A.C.M., E.R., A.P., L.G., J.H., L.B.M.K., B.W., M.H., E.P. performed experiments. F.H. designed the sgRNA library and trained the Cellpose model. M.B. acquired funding. F.P. and M.B. supervised the work. F.P. wrote the paper. All authors reviewed and edited the manuscript.

Corresponding authors

Correspondence to Fillip Port or Michael Boutros.

Competing interests

The authors declare no competing interests.

Peer review information

Nature Communications thanks Darpan Medhi and the other, anonymous, reviewer(s) for their contribution to the peer review of this work. A peer review file is available.

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