Literature DB >> 34286046

Heterologous Expression and Purification of CRISPR-Cas12a/Cpf1.

Prarthana Mohanraju1, John Van Der Oost1, Martin Jinek2, Daan C Swarts2.   

Abstract

This protocol provides step by step instructions (Figure 1) for heterologous expression of Francisella novicida Cas12a (previously known as Cpf1) in Escherichia coli. It additionally includes a protocol for high-purity purification and briefly describes how activity assays can be performed. These protocols can also be used for purification of other Cas12a homologs and the purified proteins can be used for subsequent genome editing experiments. Figure 1. Timeline of activities for the heterologous expression and purification of Francisella novicida Cas12a (FnCas12a) from Escherichia coli.
Copyright © 2018 The Authors; exclusive licensee Bio-protocol LLC.

Entities:  

Keywords:  CRISPR-Cas; Cas12a; Cpf1; Protein purification

Year:  2018        PMID: 34286046      PMCID: PMC8275269          DOI: 10.21769/BioProtoc.2842

Source DB:  PubMed          Journal:  Bio Protoc        ISSN: 2331-8325


  34 in total

1.  Use of T7 RNA polymerase to direct expression of cloned genes.

Authors:  F W Studier; A H Rosenberg; J J Dunn; J W Dubendorff
Journal:  Methods Enzymol       Date:  1990       Impact factor: 1.600

2.  Structural Basis for Guide RNA Processing and Seed-Dependent DNA Targeting by CRISPR-Cas12a.

Authors:  Daan C Swarts; John van der Oost; Martin Jinek
Journal:  Mol Cell       Date:  2017-04-20       Impact factor: 17.970

3.  Multiplex Gene Editing in Rice Using the CRISPR-Cpf1 System.

Authors:  Mugui Wang; Yanfei Mao; Yuming Lu; Xiaoping Tao; Jian-Kang Zhu
Journal:  Mol Plant       Date:  2017-03-16       Impact factor: 13.164

4.  Generation of knockout mice by Cpf1-mediated gene targeting.

Authors:  Yongsub Kim; Seung-A Cheong; Jong Geol Lee; Sang-Wook Lee; Myeong Sup Lee; In-Jeoung Baek; Young Hoon Sung
Journal:  Nat Biotechnol       Date:  2016-06-06       Impact factor: 54.908

5.  Vectors for selective expression of cloned DNAs by T7 RNA polymerase.

Authors:  A H Rosenberg; B N Lade; D S Chui; S W Lin; J J Dunn; F W Studier
Journal:  Gene       Date:  1987       Impact factor: 3.688

6.  Diversity and evolution of class 2 CRISPR-Cas systems.

Authors:  Sergey Shmakov; Aaron Smargon; David Scott; David Cox; Neena Pyzocha; Winston Yan; Omar O Abudayyeh; Jonathan S Gootenberg; Kira S Makarova; Yuri I Wolf; Konstantin Severinov; Feng Zhang; Eugene V Koonin
Journal:  Nat Rev Microbiol       Date:  2017-01-23       Impact factor: 60.633

7.  A CRISPR-Cpf1 system for efficient genome editing and transcriptional repression in plants.

Authors:  Xu Tang; Levi G Lowder; Tao Zhang; Aimee A Malzahn; Xuelian Zheng; Daniel F Voytas; Zhaohui Zhong; Yiyi Chen; Qiurong Ren; Qian Li; Elida R Kirkland; Yong Zhang; Yiping Qi
Journal:  Nat Plants       Date:  2017-02-17       Impact factor: 15.793

8.  Multiplex gene editing by CRISPR-Cpf1 using a single crRNA array.

Authors:  Bernd Zetsche; Matthias Heidenreich; Prarthana Mohanraju; Iana Fedorova; Jeroen Kneppers; Ellen M DeGennaro; Nerges Winblad; Sourav R Choudhury; Omar O Abudayyeh; Jonathan S Gootenberg; Wen Y Wu; David A Scott; Konstantin Severinov; John van der Oost; Feng Zhang
Journal:  Nat Biotechnol       Date:  2016-12-05       Impact factor: 54.908

9.  The CCTL (Cpf1-assisted Cutting and Taq DNA ligase-assisted Ligation) method for efficient editing of large DNA constructs in vitro.

Authors:  Chao Lei; Shi-Yuan Li; Jia-Kun Liu; Xuan Zheng; Guo-Ping Zhao; Jin Wang
Journal:  Nucleic Acids Res       Date:  2017-05-19       Impact factor: 16.971

10.  crRNA and tracrRNA guide Cas9-mediated DNA interference in Streptococcus thermophilus.

Authors:  Tautvydas Karvelis; Giedrius Gasiunas; Algirdas Miksys; Rodolphe Barrangou; Philippe Horvath; Virginijus Siksnys
Journal:  RNA Biol       Date:  2013-03-27       Impact factor: 4.652

View more
  2 in total

1.  Enhancement of CRISPR/Cas12a trans-cleavage activity using hairpin DNA reporters.

Authors:  Marianna Rossetti; Rosa Merlo; Neda Bagheri; Danila Moscone; Anna Valenti; Aakash Saha; Pablo R Arantes; Rudy Ippodrino; Francesco Ricci; Ida Treglia; Elisabetta Delibato; John van der Oost; Giulia Palermo; Giuseppe Perugino; Alessandro Porchetta
Journal:  Nucleic Acids Res       Date:  2022-08-12       Impact factor: 19.160

2.  CRISPR-Cas12a nucleases function with structurally engineered crRNAs: SynThetic trAcrRNA.

Authors:  D J Jedrzejczyk; L D Poulsen; M Mohr; N D Damas; S Schoffelen; A Barghetti; R Baumgartner; B T Weinert; T Warnecke; R T Gill
Journal:  Sci Rep       Date:  2022-07-16       Impact factor: 4.996

  2 in total

北京卡尤迪生物科技股份有限公司 © 2022-2023.