Animal biotechnology

  • Article
    | Open Access

    Gene drives raise safety concerns around unintended propagation. Here the authors present a trans-complementing split-gene drive that requires inheritance of separate transgenes to assemble a fully functional drive.

    • Víctor López Del Amo
    • , Alena L. Bishop
    • , Héctor M. Sánchez C.
    • , Jared B. Bennett
    • , Xuechun Feng
    • , John M. Marshall
    • , Ethan Bier
    •  & Valentino M. Gantz
  • Article
    | Open Access

    Most cases of autosomal dominant polycystic kidney disease (ADPKD) are due to mutations in PKD1. Here, Tsukiyama et al. generate monkeys with mutations in PKD1 and show that animals recapitulate key pathological features of the human disease, suggesting these may provide insights into ADPKD pathogenesis and contribute to the development of future therapeutic strategies.

    • Tomoyuki Tsukiyama
    • , Kenichi Kobayashi
    • , Masataka Nakaya
    • , Chizuru Iwatani
    • , Yasunari Seita
    • , Hideaki Tsuchiya
    • , Jun Matsushita
    • , Kahoru Kitajima
    • , Ikuo Kawamoto
    • , Takahiro Nakagawa
    • , Koji Fukuda
    • , Teppei Iwakiri
    • , Hiroyuki Izumi
    • , Iori Itagaki
    • , Shinji Kume
    • , Hiroshi Maegawa
    • , Ryuichi Nishinakamura
    • , Saori Nishio
    • , Shinichiro Nakamura
    • , Akihiro Kawauchi
    •  & Masatsugu Ema
  • Article
    | Open Access

    Multiplexed genome engineering with Cas9 can increase efficiency but also the risk of unintended alterations. Here the authors demonstrate the use of multiplexed base editors on primary T cells with reduced translocation frequency.

    • Beau R. Webber
    • , Cara-lin Lonetree
    • , Mitchell G. Kluesner
    • , Matthew J. Johnson
    • , Emily J. Pomeroy
    • , Miechaleen D. Diers
    • , Walker S. Lahr
    • , Garrett M. Draper
    • , Nicholas J. Slipek
    • , Branden A. Smeester
    • , Klaus N. Lovendahl
    • , Amber N. McElroy
    • , Wendy R. Gordon
    • , Mark J. Osborn
    •  & Branden S. Moriarity
  • Article
    | Open Access

    Lee et al. report an engineered IgG1 Fc domain that behaves like an hFcRn binding pH toggle switch. The authors show that this new half-life extension Fc domain confers improved pharmacokinetics in new humanized knock-in mouse strains that recapitulate the key processes for antibody persistence in circulation.

    • Chang-Han Lee
    • , Tae Hyun Kang
    • , Ophélie Godon
    • , Makiko Watanabe
    • , George Delidakis
    • , Caitlin M. Gillis
    • , Delphine Sterlin
    • , David Hardy
    • , Michel Cogné
    • , Lynn E. Macdonald
    • , Andrew J. Murphy
    • , Naxin Tu
    • , Jiwon Lee
    • , Jonathan R. McDaniel
    • , Emily Makowski
    • , Peter M. Tessier
    • , Aaron S. Meyer
    • , Pierre Bruhns
    •  & George Georgiou
  • Article
    | Open Access

    Selectable markers are widely used in cell engineering but there is only a limited variety to choose from. Here the authors split markers using inteins, allowing up to six transgene integration events to be selected for with one marker.

    • Nathaniel Jillette
    • , Menghan Du
    • , Jacqueline Jufen Zhu
    • , Peter Cardoz
    •  & Albert Wu Cheng
  • Article
    | Open Access

    Engineering cell lines often requires multiple plasmids with different selection markers. Here the authors present SiMPl, a method based on rationally engineered split enzymes which get reconstituted via intein-mediated protein splicing to maintain two plasmids using a single antibiotic.

    • Navaneethan Palanisamy
    • , Anna Degen
    • , Anna Morath
    • , Jara Ballestin Ballestin
    • , Claudia Juraske
    • , Mehmet Ali Öztürk
    • , Georg A. Sprenger
    • , Jung-Won Youn
    • , Wolfgang W. Schamel
    •  & Barbara Di Ventura
  • Article
    | Open Access

    DNA repair by microhomology-mediated end joining creates precise deletions based on flanking microhomologies. Here the authors use CRISPR-Cas9 to recreate pathogenic deletion mutations using existing microhomologies in the human genome identified by their program MHcut.

    • Janin Grajcarek
    • , Jean Monlong
    • , Yoko Nishinaka-Arai
    • , Michiko Nakamura
    • , Miki Nagai
    • , Shiori Matsuo
    • , David Lougheed
    • , Hidetoshi Sakurai
    • , Megumu K. Saito
    • , Guillaume Bourque
    •  & Knut Woltjen
  • Article
    | Open Access

    Previous gene editing in haematopoietic stem cells (HSCs) has focussed on a heterogeneous CD34+ population. Here, the authors demonstrate high efficiency CRISPR/Cas9-based editing of purified long-term HSCs using non-homologous end joining and homology-directed repair, by directing isoform-specific expression of GATA1.

    • Elvin Wagenblast
    • , Maria Azkanaz
    • , Sabrina A. Smith
    • , Lorien Shakib
    • , Jessica L. McLeod
    • , Gabriela Krivdova
    • , Joana Araújo
    • , Leonard D. Shultz
    • , Olga I. Gan
    • , John E. Dick
    •  & Eric R. Lechman
  • Article
    | Open Access

    The role of CTCF-bound insulator elements in enhancer-gene interactions and transcriptional regulation remains poorly understood. Here, the authors investigate multiple epigenome editing strategies for perturbing individual CTCF-bound insulators, and evaluate their effects on genome topology and transcription.

    • Daniel R. Tarjan
    • , William A. Flavahan
    •  & Bradley E. Bernstein
  • Article
    | Open Access

    Sustainable sources are needed to meet the demand for long-chain polyunsaturated fatty acids. Here the authors construct an artificial biosynthetic gene cluster in Y. lipolytica capable of producing a high concentration of PUFAs.

    • Katja Gemperlein
    • , Demian Dietrich
    • , Michael Kohlstedt
    • , Gregor Zipf
    • , Hubert S. Bernauer
    • , Christoph Wittmann
    • , Silke C. Wenzel
    •  & Rolf Müller
  • Article
    | Open Access

    Here, the authors show that reticulocytes derived from immortalized erythroblasts support invasion and development of Plasmodium falciparum and use CRISPR-mediated gene knockout and complementation of an invasion receptor to demonstrate utility of this model system for research in malaria invasion.

    • Timothy J. Satchwell
    • , Katherine E. Wright
    • , Katy L. Haydn-Smith
    • , Fernando Sánchez-Román Terán
    • , Pedro L. Moura
    • , Joseph Hawksworth
    • , Jan Frayne
    • , Ashley M. Toye
    •  & Jake Baum
  • Article
    | Open Access

    The expression of oncogenic MYC paralogs in small cell lung cancer is mutually exclusive. In this study, the authors show that MYC, but not MYCN or MYCL, represses BCL2, resulting in cells that are uniquely sensitive to apoptosis, and find that CHK1 and AURKA inhibitors may be useful for treating these cancers.

    • Marcel A. Dammert
    • , Johannes Brägelmann
    • , Rachelle R. Olsen
    • , Stefanie Böhm
    • , Niloufar Monhasery
    • , Christopher P. Whitney
    • , Milind D. Chalishazar
    • , Hannah L. Tumbrink
    • , Matthew R. Guthrie
    • , Sebastian Klein
    • , Abbie S. Ireland
    • , Jeremy Ryan
    • , Anna Schmitt
    • , Annika Marx
    • , Luka Ozretić
    • , Roberta Castiglione
    • , Carina Lorenz
    • , Ron D. Jachimowicz
    • , Elmar Wolf
    • , Roman K. Thomas
    • , John T. Poirier
    • , Reinhard Büttner
    • , Triparna Sen
    • , Lauren A. Byers
    • , H. Christian Reinhardt
    • , Anthony Letai
    • , Trudy G. Oliver
    •  & Martin L. Sos
  • Article
    | Open Access

    Manipulating DNA repair pathways can be used to improve the outcomes of CRISPR-based genome editing. Here the authors derive an enhanced RAD18 variant that suppresses 53BP1 recruitment to DNA double-strand breaks to enhance homology-mediated repair.

    • Tarun S. Nambiar
    • , Pierre Billon
    • , Giacomo Diedenhofen
    • , Samuel B. Hayward
    • , Angelo Taglialatela
    • , Kunheng Cai
    • , Jen-Wei Huang
    • , Giuseppe Leuzzi
    • , Raquel Cuella-Martin
    • , Andrew Palacios
    • , Anuj Gupta
    • , Dieter Egli
    •  & Alberto Ciccia
  • Article
    | Open Access

    Construction of plasmids from multiple fragments often uses customised parts and leaves scars where fragments are joined. Here the authors develop a method for barcoding fragments and constructing plasmids in a scarless manner from a collection of standard parts.

    • Xiaoqiang Ma
    • , Hong Liang
    • , Xiaoyi Cui
    • , Yurou Liu
    • , Hongyuan Lu
    • , Wenbo Ning
    • , Nga Yu Poon
    • , Benjamin Ho
    •  & Kang Zhou
  • Article
    | Open Access

    Inefficient delivery of proteins into mammalian cells limits their use in research and therapy. Here, the authors discover that ProTα, a small, intrinsically disordered human protein can facilitate efficient cationic lipid-mediated protein delivery of genome editing proteins into mammalian cells.

    • Y. Bill Kim
    • , Kevin T. Zhao
    • , David B. Thompson
    •  & David R. Liu
  • Article
    | Open Access

    Studies with patient derived xenografts are hampered by factors such as genetic variability and sample availability. Here, the authors generate a leukemia mouse model by lentiviral transduction of normal human cord blood and show an oncogenic role of HOXB genes.

    • Manabu Kusakabe
    • , Ann Chong Sun
    • , Kateryna Tyshchenko
    • , Rachel Wong
    • , Aastha Nanda
    • , Claire Shanna
    • , Samuel Gusscott
    • , Elizabeth A. Chavez
    • , Alireza Lorzadeh
    • , Alice Zhu
    • , Ainsleigh Hill
    • , Stacy Hung
    • , Scott Brown
    • , Artem Babaian
    • , Xuehai Wang
    • , Robert A. Holt
    • , Christian Steidl
    • , Aly Karsan
    • , R. Keith Humphries
    • , Connie J. Eaves
    • , Martin Hirst
    •  & Andrew P. Weng
  • Article
    | Open Access

    Synthetic promoters can be superior to native ones but the design is challenging without knowledge of gene regulation. Here the authors develop a pipeline that allows for screening a synthetic promoter library to identify high performance promoters in potentially any given cell state of interest.

    • Ming-Ru Wu
    • , Lior Nissim
    • , Doron Stupp
    • , Erez Pery
    • , Adina Binder-Nissim
    • , Karen Weisinger
    • , Casper Enghuus
    • , Sebastian R. Palacios
    • , Melissa Humphrey
    • , Zhizhuo Zhang
    • , Eva Maria Novoa
    • , Manolis Kellis
    • , Ron Weiss
    • , Samuel D. Rabkin
    • , Yuval Tabach
    •  & Timothy K. Lu
  • Article
    | Open Access

    Roughly 25% of mouse genes are embryonically lethal when knocked out, preventing the generation of viable mouse models. Here the authors use CRISPR-Cas9 to edit one blastomere of a two-cell embryo to generate viable chimeric mice.

    • Yi Wu
    • , Jing Zhang
    • , Boya Peng
    • , Dan Tian
    • , Dong Zhang
    • , Yang Li
    • , Xiaoyu Feng
    • , Jinghao Liu
    • , Jun Li
    • , Teng Zhang
    • , Xiaoyong Liu
    • , Jing Lu
    • , Baian Chen
    •  & Songlin Wang
  • Article
    | Open Access

    Base editors can efficiently produce single nucleotide alterations without requiring a double-strand break. Here the authors show base editing at multiple sites simultaneously in pigs.

    • Jingke Xie
    • , Weikai Ge
    • , Nan Li
    • , Qishuai Liu
    • , Fangbing Chen
    • , Xiaoyu Yang
    • , Xingyun Huang
    • , Zhen Ouyang
    • , Quanjun Zhang
    • , Yu Zhao
    • , Zhaoming Liu
    • , Shixue Gou
    • , Han Wu
    • , Chengdan Lai
    • , Nana Fan
    • , Qin Jin
    • , Hui Shi
    • , Yanhui Liang
    • , Ting Lan
    • , Longquan Quan
    • , Xiaoping Li
    • , Kepin Wang
    •  & Liangxue Lai
  • Article
    | Open Access

    Global inhibition of NHEJ factors has been one strategy to improve CRISPR-Cas9 mediated HDR. Here the authors fuse a dominant-negative mutant of 53BP1 to Cas9 to enhance HDR frequency, reduce NHEJ specifically at the Cas9 cut sites, and reduce the toxicity associated with global NHEJ inhibition.

    • Rajeswari Jayavaradhan
    • , Devin M. Pillis
    • , Michael Goodman
    • , Fan Zhang
    • , Yue Zhang
    • , Paul R. Andreassen
    •  & Punam Malik
  • Article
    | Open Access

    Here the authors fuse hRad51 and variants thereof to Cas9 nickase to facilitate homology-directed repair without generating double strand breaks, minimizing indel formation and off-target editing. This tool represents progress towards the goal of performing HDR without an excess of undesired side products.

    • Holly A. Rees
    • , Wei-Hsi Yeh
    •  & David R. Liu
  • Article
    | Open Access

    The development of site-specific recombinases as genome editing tools is limited by the difficulty of altering their DNA sequence specificity. Here the authors present Rec-seq, a method for identifying specificity determinants and off-target substrates of recombinases in an unbiased manner.

    • Jeffrey L. Bessen
    • , Lena K. Afeyan
    • , Vlado Dančík
    • , Luke W. Koblan
    • , David B. Thompson
    • , Chas Leichner
    • , Paul A. Clemons
    •  & David R. Liu
  • Review Article
    | Open Access

    Generation of transgenic mice has become routine in studying gene function and disease mechanisms, but often this is not enough to fully understand human biology. Here, the authors review the current state of the art of targeted genomic humanisation strategies and their advantages over classic approaches.

    • Fei Zhu
    • , Remya R. Nair
    • , Elizabeth M. C. Fisher
    •  & Thomas J. Cunningham
  • Article
    | Open Access

    Gene-drives use CRISPR-Cas9 to be transmitted in a super-Mendelian fashion. Here the authors develop an allelic-drive for selective inheritance of a desired allele.

    • Annabel Guichard
    • , Tisha Haque
    • , Marketta Bobik
    • , Xiang-Ru S. Xu
    • , Carissa Klanseck
    • , Raja Babu Singh Kushwah
    • , Mateus Berni
    • , Bhagyashree Kaduskar
    • , Valentino M. Gantz
    •  & Ethan Bier
  • Article
    | Open Access

    Gene correction in hematopoietic stem cells could be a powerful way to treat monogenic diseases of the blood and immune system. Here the authors develop a strategy using CRISPR-Cas9 and an aAdeno-Associated vVirus(AAV)-delivered IL2RG cDNA to correct X-linked sSevere Ccombined iImmunodeficiency (SCID-X1) with a high success rate.

    • Mara Pavel-Dinu
    • , Volker Wiebking
    • , Beruh T. Dejene
    • , Waracharee Srifa
    • , Sruthi Mantri
    • , Carmencita E. Nicolas
    • , Ciaran Lee
    • , Gang Bao
    • , Eric J. Kildebeck
    • , Niraj Punjya
    • , Camille Sindhu
    • , Matthew A. Inlay
    • , Nivedita Saxena
    • , Suk See DeRavin
    • , Harry Malech
    • , Maria Grazia Roncarolo
    • , Kenneth I. Weinberg
    •  & Matthew H. Porteus
  • Article
    | Open Access

    A desired product cannot be obtained at higher concentration than its equilibrium concentration when isomerases are used for biotransformation. Here, the authors engineer in vivo oxidoreductive reactions in yeast to overcome the equilibrium limitation of in vitro isomerases-based tagatose production.

    • Jing-Jing Liu
    • , Guo-Chang Zhang
    • , Suryang Kwak
    • , Eun Joong Oh
    • , Eun Ju Yun
    • , Kulika Chomvong
    • , Jamie H. D. Cate
    •  & Yong-Su Jin
  • Article
    | Open Access

    Mutations that modulate the activity of ion channels are essential tools to understand the biophysical determinants that control their gating. Here authors reveal the role played by a single residue in the second transmembrane domain of vertebrate and invertebrate two-pore domain potassium channels.

    • Ismail Ben Soussia
    • , Sonia El Mouridi
    • , Dawon Kang
    • , Alice Leclercq-Blondel
    • , Lamyaa Khoubza
    • , Philippe Tardy
    • , Nora Zariohi
    • , Marie Gendrel
    • , Florian Lesage
    • , Eun-Jin Kim
    • , Delphine Bichet
    • , Olga Andrini
    •  & Thomas Boulin
  • Article
    | Open Access

    It is difficult to identify cancer driver genes in cancers, for instance BRCA1 mutated breast cancer, that are characterised by large scale genomic alterations. Here, the authors develop genetically engineered mouse models of BRCA1-deficient breast cancer that allow highthroughput in vivo perturbation of candidate driver genes, validating drivers Myc, Met, Pten and Rb1, and identifying MCL1 as a collaborating driver whose targeting can impact efficacy of PARP inhibition.

    • Stefano Annunziato
    • , Julian R. de Ruiter
    • , Linda Henneman
    • , Chiara S. Brambillasca
    • , Catrin Lutz
    • , François Vaillant
    • , Federica Ferrante
    • , Anne Paulien Drenth
    • , Eline van der Burg
    • , Bjørn Siteur
    • , Bas van Gerwen
    • , Roebi de Bruijn
    • , Martine H. van Miltenburg
    • , Ivo J. Huijbers
    • , Marieke van de Ven
    • , Jane E. Visvader
    • , Geoffrey J. Lindeman
    • , Lodewyk F. A. Wessels
    •  & Jos Jonkers
  • Article
    | Open Access

    Most approaches to control gene expression in vivo require generation of knock-in mouse lines and often lack spatiotemporal control. Here the authors develop a photo-activatable Flp recombinase system and demonstrate its use by controlling object-exploration behavior in mice through Cav3.1 silencing.

    • Hyunjin Jung
    • , Seong-Wook Kim
    • , Minsoo Kim
    • , Jongryul Hong
    • , Daseuli Yu
    • , Ji Hye Kim
    • , Yunju Lee
    • , Sungsoo Kim
    • , Doyeon Woo
    • , Hee-Sup Shin
    • , Byung Ouk Park
    •  & Won Do Heo
  • Article
    | Open Access

    A current challenge in genome editing is delivering Cas9 and sgRNA into target cells. Here the authors engineer a delivery system based on murine leukemia virus-like particles loaded with Cas9-sgRNA ribonucleoproteins to induce efficient genome editing in both cell culture and in vivo in mouse.

    • Philippe E. Mangeot
    • , Valérie Risson
    • , Floriane Fusil
    • , Aline Marnef
    • , Emilie Laurent
    • , Juliana Blin
    • , Virginie Mournetas
    • , Emmanuelle Massouridès
    • , Thibault J. M. Sohier
    • , Antoine Corbin
    • , Fabien Aubé
    • , Marie Teixeira
    • , Christian Pinset
    • , Laurent Schaeffer
    • , Gaëlle Legube
    • , François-Loïc Cosset
    • , Els Verhoeyen
    • , Théophile Ohlmann
    •  & Emiliano P. Ricci
  • Article
    | Open Access

    Therapeutic genome engineering relies on the development of reliable, robust and versatile tools. Here the authors develop Cas9-Cas9 chimeras with high target site activity that generate predictable deletions.

    • Mehmet Fatih Bolukbasi
    • , Pengpeng Liu
    • , Kevin Luk
    • , Samantha F. Kwok
    • , Ankit Gupta
    • , Nadia Amrani
    • , Erik J. Sontheimer
    • , Lihua Julie Zhu
    •  & Scot A. Wolfe
  • Article
    | Open Access

    Understanding the risks of bystander and off-target editing is essential for genome engineering applications. Here, the authors analyze the fidelity of adenine and cytosine base editors in vivo.

    • Hye Kyung Lee
    • , Michaela Willi
    • , Shannon M. Miller
    • , Sojung Kim
    • , Chengyu Liu
    • , David R. Liu
    •  & Lothar Hennighausen
  • Article
    | Open Access

    Vector alteration strategies are emerging as attractive tools for malaria transmission control. Here, Shane et al. engineer a bacterial strain, isolated from mosquitoes, to produce an antiplasmodial protein in the presence of blood meal, causing the mosquitoes to become refractory to Plasmodium infection.

    • Jackie L. Shane
    • , Christina L. Grogan
    • , Caroline Cwalina
    •  & David J. Lampe
  • Article
    | Open Access

    A bottleneck for the application of bioelectrochemical systems is the slow rate of extracellular electron transfer. Here the authors use a synthetic biology approach to redirect metabolic flux to NAD+ biosynthesis, which enhances the intracellular electron flux and the extracellular electron transfer rate.

    • Feng Li
    • , Yuan-Xiu Li
    • , Ying-Xiu Cao
    • , Lei Wang
    • , Chen-Guang Liu
    • , Liang Shi
    •  & Hao Song
  • Article
    | Open Access

    Current approaches to conditionally deplete target proteins require site-specific genetic engineering or have poor temporal control. Here the authors overcome these limitations by combining the AID system with nanobodies to reversibly degrade GFP-tagged proteins in living cells and zebrafish.

    • Katrin Daniel
    • , Jaroslav Icha
    • , Cindy Horenburg
    • , Doris Müller
    • , Caren Norden
    •  & Jörg Mansfeld
  • Article
    | Open Access

    Optimization of the recently discovered Class 2 CRISPR protein Cpf1 has the potential to promote its applications in gene editing and therapeutics. Here, the authors find that extending the 5′ end of the crRNA can increase both the editing efficiency and delivery of Cpf1 in vitro and in vivo.

    • Hyo Min Park
    • , Hui Liu
    • , Joann Wu
    • , Anthony Chong
    • , Vanessa Mackley
    • , Christof Fellmann
    • , Anirudh Rao
    • , Fuguo Jiang
    • , Hunghao Chu
    • , Niren Murthy
    •  & Kunwoo Lee
  • Article
    | Open Access

    Genome editing using CRISPR can be enhanced by manipulating DNA double-strand break repair pathways. Here the authors demonstrate LoAD, local accumulation of repair molecules, which shifts repair to microhomology-mediated end-joining.

    • Shota Nakade
    • , Keiji Mochida
    • , Atsushi Kunii
    • , Kazuki Nakamae
    • , Tomomi Aida
    • , Kohichi Tanaka
    • , Naoaki Sakamoto
    • , Tetsushi Sakuma
    •  & Takashi Yamamoto
  • Article
    | Open Access

    Delivery of Cas9 ribonucleoprotein complexes can be done via microinjection into eggs, though this is a technically challenging procedure. Here the authors demonstrate ReMOT Control, delivery of RNPs to the mosquito germline by tagging with peptide derived from yolk protein 1.

    • Duverney Chaverra-Rodriguez
    • , Vanessa M. Macias
    • , Grant L. Hughes
    • , Sujit Pujhari
    • , Yasutsugu Suzuki
    • , David R. Peterson
    • , Donghun Kim
    • , Sage McKeand
    •  & Jason L. Rasgon
  • Article
    | Open Access

    A rigorous understanding of off-target effects is necessary for SaCas9 to be used in therapeutic genome editing. Here the authors measure SaCas9 mismatch tolerance across a pairwise library screen of 88,000 guides and targets in human cells and develop a model which ranks off-target sites.

    • Josh Tycko
    • , Luis A. Barrera
    • , Nicholas C. Huston
    • , Ari E. Friedland
    • , Xuebing Wu
    • , Jonathan S. Gootenberg
    • , Omar O. Abudayyeh
    • , Vic E. Myer
    • , Christopher J. Wilson
    •  & Patrick D. Hsu
  • Article
    | Open Access

    The correction of genetic defects in utero could allow for improved outcomes of gene therapy. Here, the authors demonstrate safe delivery of nanoparticles to fetal mouse tissues, and show that nanoparticles containing peptide nucleic acids to edit the beta-globin gene are effective in a mouse model of beta-thalassemia.

    • Adele S. Ricciardi
    • , Raman Bahal
    • , James S. Farrelly
    • , Elias Quijano
    • , Anthony H. Bianchi
    • , Valerie L. Luks
    • , Rachael Putman
    • , Francesc López-Giráldez
    • , Süleyman Coşkun
    • , Eric Song
    • , Yanfeng Liu
    • , Wei-Che Hsieh
    • , Danith H. Ly
    • , David H. Stitelman
    • , Peter M. Glazer
    •  & W. Mark Saltzman
  • Article
    | Open Access

    CRISPR-based base editors allow for single nucleotide genome editing in a range of organisms. Here the authors demonstrate the in vivo generation of mouse models carrying clinically relevant mutations using C→T and A→G editors.

    • Zhen Liu
    • , Zongyang Lu
    • , Guang Yang
    • , Shisheng Huang
    • , Guanglei Li
    • , Songjie Feng
    • , Yajing Liu
    • , Jianan Li
    • , Wenxia Yu
    • , Yu Zhang
    • , Jia Chen
    • , Qiang Sun
    •  & Xingxu Huang
  • Article
    | Open Access

    Base editing allows the precise introduction of point mutations into cellular DNA without requiring double-stranded DNA breaks or homology-directed repair, which is inefficient in postmitotic cells. Here the authors demonstrate in vivo base editing of post-mitotic somatic cells in the postnatal mouse inner ear with physiological outcomes.

    • Wei-Hsi Yeh
    • , Hao Chiang
    • , Holly A. Rees
    • , Albert S. B. Edge
    •  & David R. Liu
  • Article
    | Open Access

    The International Synthetic Yeast Sc2.0 project has built Cre recombinase sites into synthetic chromosomes, enabling rapid genome evolution. Here the authors demonstrate L-SCRaMbLE, a light-controlled recombinase tool with improved control over recombination events.

    • Lena Hochrein
    • , Leslie A. Mitchell
    • , Karina Schulz
    • , Katrin Messerschmidt
    •  & Bernd Mueller-Roeber
  • Article
    | Open Access

    DNA double-strand break (DSB) leads to genome rearrangements with various genetic and phenotypic effects. Here, the authors develop a tool to induce large-scale genome restructuring by introducing conditional multiple DNA breaks, and produce various traits in yeast and Arabidopsis thaliana.

    • Nobuhiko Muramoto
    • , Arisa Oda
    • , Hidenori Tanaka
    • , Takahiro Nakamura
    • , Kazuto Kugou
    • , Kazuki Suda
    • , Aki Kobayashi
    • , Shiori Yoneda
    • , Akinori Ikeuchi
    • , Hiroki Sugimoto
    • , Satoshi Kondo
    • , Chikara Ohto
    • , Takehiko Shibata
    • , Norihiro Mitsukawa
    •  & Kunihiro Ohta
  • Article
    | Open Access

    Efficient gene targeting in higher plants remains challenging. Here, the authors develop a sequential transformation method for CRISPR/Cas9-mediated gene targeting in Arabidopsis and demonstrate its functionality at five genomic sites in two endogenous loci.

    • Daisuke Miki
    • , Wenxin Zhang
    • , Wenjie Zeng
    • , Zhengyan Feng
    •  & Jian-Kang Zhu