TALEN or Cas9 – Rapid, Efficient and Specific Choices for Genome Modifications

Wei, Chuanxian; Liu, Jiyong; Yu, Zhongsheng; Zhang, Bo; Gao, Guanjun; Jiao, Renjie

doi:10.1016/j.jgg.2013.03.013

Cited by 147 publications

(106 citation statements)

References 90 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…DSBs made by any site-specific nucleases should possess the capacity to enter the homology-directed repair pathway and generate precisely specified changes in the genome. Effective nucleases with engineered specificity include the zinc-finger nucleases (ZFNs), the transcription activatorlike effector (TALE) nucleases (TALENs), and the RNAguided CRISPR-associated (Cas9) endonuclease (Urnov et al 2010;Bogdanove and Voytas 2011;Jinek et al 2012;Wiedenheft et al 2012;Gaj et al 2013;Wei et al 2013). ZFNs and TALENs contain fusions between the DNA cleavage domain of the FokI endonuclease and a customdesigned DNA-binding domain: either C 2 H 2 zinc-finger motifs for ZFNs or TALE domains for TALENs (Urnov et al 2010;Carroll 2011;Cermak et al 2011;Li et al 2011;Miller et al 2011;Mussolino et al 2011).…”

mentioning

confidence: 99%

Precise and Heritable Genome Editing in Evolutionarily Diverse Nematodes Using TALENs and CRISPR/Cas9 to Engineer Insertions and Deletions

et al. 2013

View full text Add to dashboard Cite

Exploitation of custom-designed nucleases to induce DNA double-strand breaks (DSBs) at genomic locations of choice has transformed our ability to edit genomes, regardless of their complexity. DSBs can trigger either error-prone repair pathways that induce random mutations at the break sites or precise homology-directed repair pathways that generate specific insertions or deletions guided by exogenously supplied DNA. Prior editing strategies using site-specific nucleases to modify the Caenorhabditis elegans genome achieved only the heritable disruption of endogenous loci through random mutagenesis by error-prone repair. Here we report highly effective strategies using TALE nucleases and RNA-guided CRISPR/Cas9 nucleases to induce error-prone repair and homology-directed repair to create heritable, precise insertion, deletion, or substitution of specific DNA sequences at targeted endogenous loci. Our robust strategies are effective across nematode species diverged by 300 million years, including necromenic nematodes (Pristionchus pacificus), male/female species (Caenorhabditis species 9), and hermaphroditic species (C. elegans). Thus, genome-editing tools now exist to transform nonmodel nematode species into genetically tractable model organisms. We demonstrate the utility of our broadly applicable genome-editing strategies by creating reagents generally useful to the nematode community and reagents specifically designed to explore the mechanism and evolution of X chromosome dosage compensation. By developing an efficient pipeline involving germline injection of nuclease mRNAs and single-stranded DNA templates, we engineered precise, heritable nucleotide changes both close to and far from DSBs to gain or lose genetic function, to tag proteins made from endogenous genes, and to excise entire loci through targeted FLP-FRT recombination. STRATEGIES to engineer heritable, site-directed mutations at endogenous loci have revolutionized our approach toward manipulating and dissecting genome function. Studies of plants and animals alike, whether conducted in whole organisms or cell lines, have benefitted greatly from these genomeediting approaches (Bibikova et al. 2002;Beumer et al. 2006;Doyon et al. 2008;Geurts et al. 2009;Hockemeyer et al. 2009;Holt et al. 2010;Zhang et al. 2010;Hockemeyer et al. 2011;Tesson et al. 2011;Wood et al. 2011;Young et al. 2011;Bedell et al. 2012;Bassett et al. 2013;Cong et al. 2013;Jinek et al. 2013;Mali et al. 2013;Wang et al. 2013;Zu et al. 2013). The most modern tools for modifying complex genomes at single-nucleotide resolution are site-specific nucleases that induce DNA double-strand breaks (DSBs) at specifically designated genomic locations. DSBs trigger repair pathways that can elicit targeted genetic reprogramming, primarily through two mechanisms: error-prone nonhomologous end joining (NHEJ) (Lieber 2010) and precise, homology-directed recombination or repair (HDR) (Chapman et al. 2012). NHEJ rejoins broken ends of chromosomes through an imprecise process that produces nucleo...

show abstract

mentioning

confidence: 99%

Precise and Heritable Genome Editing in Evolutionarily Diverse Nematodes Using TALENs and CRISPR/Cas9 to Engineer Insertions and Deletions

et al. 2013

View full text Add to dashboard Cite

show abstract

“…[15] and [16]). The mechanism is efficient in dividing cells and has been widely exploited for insertion of sequences at particular target sites.…”

Section: Discussionmentioning

confidence: 98%

“…[15] and [16]). HDR is based on producing DSBs at specific target sites and simultaneous introduction of 'repairing' donor DNA.…”

Section: Introductionmentioning

confidence: 98%

“…The principle underlying introduction of pri-miR-31/5/8/9 sequences into HBV DNA is based on improved efficiency of HDR in dividing cells following formation of DSBs in the target DNA (reviewed in Refs. [15] and [16]). In our scheme, the process is initiated by cleavage of HBV DNA by the TALENs (Fig.…”

Section: Using Talens To Introduce Artificial Anti-hbv Pri-mirs Into mentioning

confidence: 98%

See 1 more Smart Citation

Improved antiviral efficacy using TALEN-mediated homology directed recombination to introduce artificial primary miRNAs into DNA of hepatitis B virus

Dreyer

Nicholson

Ely

et al. 2016

Biochemical and Biophysical Research Communications

View full text Add to dashboard Cite

Highlights• Chronic infection with HBV is attributed to cccDNA persistence.• TALENs and miRNA activators have shown anti-HBV therapeutic promise.• HDR of HBV targets with TALENs and pri-miRNA donors augmented antiviral efficacy.• Combining gene editing and silencing improves inhibition of HBV gene expression. AbstractChronic infection with hepatitis B virus (HBV) remains an important global health problem. Currently licensed therapies have modest curative efficacy, which is as a result of their transient effects and limited action on the viral replication intermediate comprising covalently closed circular DNA (cccDNA). Gene editing with artificial HBV-specific endonucleases and use of artificial activators of the RNA interference pathway have shown anti-HBV therapeutic promise. Although results from these gene therapies are encouraging, maximizing durable antiviral effects is important. To address this goal, a strategy that entails combining gene editing with homology-directed DNA recombination (HDR), to introduce HBV-silencing artificial primary microRNAs (pri-miRs) into HBV DNA targets, is reported here. Previously described transcription activator-like effector nucleases (TALENs) that target the core and surface sequences of HBV were used to introduce double stranded breaks in the viral DNA. Simultaneous administration of donor sequences encoding artificial promoterless anti-HBV pri-miRs, with flanking arms that were homologous to sequences adjoining the TALENs' targets, augmented antiviral efficacy. Analysis showed targeted integration and the length of the flanking homologous arms of donor DNA had a minimal effect on antiviral efficiency. These results support the notion that gene editing and silencing may be combined to effect improved inhibition of HBV gene expression.

show abstract

“…In addition, pigs generated through SCNT often show developmental defects, which is the reason for the high mortality rate at birth observed in HCKO pigs in this study (data not shown) and other studies with genetically modified pigs using SCNT. However, in recent years, the explosive development in porcine gene-editing technologies based on meganucleases (Zinc Finger Nucleases, Tal effector nucleases, CRISPR/Cas9) has provided new tools that may help to get around the need for SCNT for developing genetically modified pigs with a much higher efficiency at lower costs (16,28,44). Genetically modified pigs are becoming more readily available for the study of mechanisms of protective immunity against human viral diseases.…”

Section: Discussionmentioning

confidence: 99%

B-Cell-Deficient and CD8 T-Cell-Depleted Gnotobiotic Pigs for the Study of Human Rotavirus Vaccine-Induced Protective Immune Responses

et al. 2016

View full text Add to dashboard Cite

Genetically modified pigs have become available recently. In this study, we established the gnotobiotic pig model of human rotavirus (HRV) infection using cloned pigs with homozygous disruption in the gene encoding immunoglobulin heavy chain (HCKO), which totally impairs B-cell development. To clarify importance of B cells and cytotoxic T cells in rotavirus immunity, CD8 cells in a subset of the pigs were depleted by injecting antipig CD8 antibodies and the immune phenotypes of all pigs were examined. HCKO pigs, CD8 cell-depleted HCKO pigs, and wild-type (WT) pigs were vaccinated with an attenuated HRV vaccine and challenged with virulent HRV. Protection against HRV infection and diarrhea was assessed postchallenge and detailed T-cell subset responses were determined pre-and postchallenge. Significantly longer duration of virus shedding was seen in vaccinated HCKO pigs than in WT pigs, indicating the importance of B cells in vaccine-induced protective immunity. Vaccinated HCKO/CD8-pigs shed significantly higher number of infectious virus than WT pigs and non-CD8-depleted HCKO pigs, indicating the importance of CD8 T cells in controlling virus replication. Therefore, both B cells and CD8 T cells play an important role in the protection against rotavirus infection. HCKO and HCKO/CD8-pigs did not differ significantly in diarrhea and virus shedding postchallenge; increased CD4 and CD8-cd T-cell responses probably compensated partially for the lack of CD8 T cells. This study demonstrated that HCKO pigs can serve as a valuable model for dissection of protective immune responses against viral infections and diseases.

show abstract

TALEN or Cas9 – Rapid, Efficient and Specific Choices for Genome Modifications

Cited by 147 publications

References 90 publications

Precise and Heritable Genome Editing in Evolutionarily Diverse Nematodes Using TALENs and CRISPR/Cas9 to Engineer Insertions and Deletions

Precise and Heritable Genome Editing in Evolutionarily Diverse Nematodes Using TALENs and CRISPR/Cas9 to Engineer Insertions and Deletions

Improved antiviral efficacy using TALEN-mediated homology directed recombination to introduce artificial primary miRNAs into DNA of hepatitis B virus

B-Cell-Deficient and CD8 T-Cell-Depleted Gnotobiotic Pigs for the Study of Human Rotavirus Vaccine-Induced Protective Immune Responses

Contact Info

Product

Resources

About