J. Keith Joung
Harvard University
Genome editingMolecular biologyZinc fingerGeneticsBiology
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Publications 166
#1Lilian Cruz (Harvard University)H-Index: 2
#2Bence GyörgyH-Index: 20
Last. J. Keith Joung (Harvard University)H-Index: 62
view all 13 authors...
Abstract Most individuals affected with DYT1 dystonia have a heterozygous three base-pair deletion in the TOR1A gene (c.907_909delGAG). The mutation appears to act through a dominant negative mechanism compromising normal torsinA function, and it is proposed that reducing mutant torsinA may normalize torsinA activity. Here we used an engineered Cas9 variant from Streptococcus pyogenes (SpCas9-VRQR) to target the mutation in TOR1A gene in order to disrupt mutant torsinA in DYT1 patient fibroblast...
#1Jonathan Y. Hsu (MIT: Massachusetts Institute of Technology)H-Index: 6
#2Andrew V. Anzalone (Broad Institute)
Last. Luca Pinello (Harvard University)H-Index: 22
view all 8 authors...
Prime editing (PE) is a versatile genome editing technology, but design of the required guide RNAs is more complex than for standard CRISPR-based nucleases or base editors. Here we describe PrimeDesign, a user-friendly, end-to-end web application and command-line tool for the design of PE experiments. PrimeDesign can be used for single and combination editing applications, as well as genome-wide and saturation mutagenesis screens. Using PrimeDesign, we also constructed PrimeVar, the first compre...
#1Jing Zeng (Harvard University)H-Index: 1
#2Yuxuan Wu (ECNU: East China Normal University)H-Index: 4
Last. Qiuming Yao (Harvard University)H-Index: 9
view all 16 authors...
Base editing by nucleotide deaminases linked to programmable DNA-binding proteins represents a promising approach to permanently remedy blood disorders, although its application in engrafting hematopoietic stem cells (HSCs) remains unexplored. In this study, we purified A3A (N57Q)-BE3 base editor for ribonucleoprotein (RNP) electroporation of human-peripheral-blood-mobilized CD34+ hematopoietic stem and progenitor cells (HSPCs). We observed frequent on-target cytosine base edits at the BCL11A er...
1 CitationsSource
#1M. Ma (Georgia Regents University)H-Index: 1
#1Manxiu Ma (Georgia Regents University)H-Index: 1
Last. Y. Albert PanH-Index: 9
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Down Syndrome Cell Adhesion Molecules (dscam and dscaml1) are essential regulators of neural circuit assembly, but their roles in vertebrate neural circuit function are still mostly unexplored. We investigated the functional consequences of dscaml1 deficiency in the larval zebrafish (sexually undifferentiated) oculomotor system, where behavior, circuit function, and neuronal activity can be precisely quantified. Genetic perturbation of dscaml1 resulted in deficits in retinal patterning and light...
1 CitationsSource
#1Killian S. Hanlon (Harvard University)H-Index: 1
#2Benjamin P. Kleinstiver (Harvard University)H-Index: 15
Last. Bence György (Harvard University)H-Index: 20
view all 19 authors...
Adeno-associated virus (AAV) vectors have shown promising results in preclinical models, but the genomic consequences of transduction with AAV vectors encoding CRISPR-Cas nucleases is still being examined. In this study, we observe high levels of AAV integration (up to 47%) into Cas9-induced double-strand breaks (DSBs) in therapeutically relevant genes in cultured murine neurons, mouse brain, muscle and cochlea. Genome-wide AAV mapping in mouse brain shows no overall increase of AAV integration ...
8 CitationsSource
#1Julian Grünewald (Harvard University)H-Index: 2
#1Julian Grünewald (Harvard University)H-Index: 1
Last. J. Keith Joung (Harvard University)H-Index: 62
view all 7 authors...
Cytosine or adenine base editors (CBEs or ABEs) can introduce specific DNA C-to-T or A-to-G alterations1–4. However, we recently demonstrated that they can also induce transcriptome-wide guide-RNA-independent editing of RNA bases5, and created selective curbing of unwanted RNA editing (SECURE)-BE3 variants that have reduced unwanted RNA-editing activity5. Here we describe structure-guided engineering of SECURE-ABE variants with reduced off-target RNA-editing activity and comparable on-target DNA...
14 CitationsSource
#1Kendall R Sanson (Broad Institute)H-Index: 3
#2Peter C DeWeirdt (Broad Institute)H-Index: 1
Last. John G. Doench (Broad Institute)H-Index: 29
view all 13 authors...
Cas12a enzymes have attractive properties for scalable delivery of multiplexed perturbations, yet widespread usage has lagged behind Cas9-based strategies. Here we describe the optimization of Cas12a from Acidaminococcus (AsCas12a) for use in pooled genetic screens in human cells. By assaying the activity of thousands of guides, we confirm on-target design rules and extend them to an enhanced activity variant, enAsCas12a. We also develop the first comprehensive set of off-target rules for Cas12a...
1 CitationsSource
#1Bence György (Harvard University)H-Index: 20
#2Carl Nist-Lund (Boston Children's Hospital)H-Index: 3
Last. David P. Corey (Harvard University)H-Index: 75
view all 15 authors...
Since most dominant human mutations are single nucleotide substitutions1,2, we explored gene editing strategies to disrupt dominant mutations efficiently and selectively without affecting wild-type alleles. However, single nucleotide discrimination can be difficult to achieve3 because commonly used endonucleases, such as Streptococcus pyogenes Cas9 (SpCas9), can tolerate up to seven mismatches between guide RNA (gRNA) and target DNA. Furthermore, the protospacer-adjacent motif (PAM) in some Cas9...
6 CitationsSource
#1Julian Grünewald (Harvard University)H-Index: 2
#2Ronghao Zhou (Harvard University)H-Index: 2
Last. J. Keith Joung (Harvard University)H-Index: 62
view all 7 authors...
Abstract CRISPR-guided DNA base editors enable the efficient installation of targeted single-nucleotide changes. Cytosine or adenine base editors (CBEs or ABEs), which are fusions of cytidine or adenosine deaminases to CRISPR-Cas nickases, can efficiently induce DNA C-to-T or A-to-G alterations in DNA, respectively1-4. We recently demonstrated that both the widely used CBE BE3 (harboring a rat APOBEC1 cytidine deaminase) and the optimized ABEmax editor can induce tens of thousands of guide RNA-i...
3 CitationsSource
#1Eric S. LanderH-Index: 252
#2Françoise BaylisH-Index: 24
Last. Ernst-Ludwig WinnackerH-Index: 1
view all 18 authors...
Eric Lander, Francoise Baylis, Feng Zhang, Emmanuelle Charpentier, Paul Berg and specialists from seven countries call for an international governance framework. Eric Lander, Francoise Baylis, Feng Zhang, Emmanuelle Charpentier, Paul Berg and specialists from seven countries call for an international governance framework.
31 CitationsSource