Using figure 1 and the following background information answer the following questions. Identification of the genetic cause of hornlessness in cattle has been the subject of intensive genetic and genomic research, culminating in the nomination of two different candidate neomutations on cattle chromosome 1 that are predicted to have arisen 500-1,000 years ago: a complex allele of Friesian origin (PF), an 80,128 base pair (bp) duplication (1909352–1989480 bp), and a second, simple allele of Celtic origin (PC) corresponding to a duplication of 212 bp (chromosome 1 positions 1705834–1706045) in place of a 10-bp deletion (1706051–1706060)We report the use of genome editing using transcription activator-like effector nucleases (TALENs) to introgress the putative PC POLLED allele into the genome of bovine embryo fibroblasts to try and produce a genotype identical to what is achievable using natural mating, but without the attendant genetic drag and admixture. In our previous studies, we used TALEN-stimulated homology-dependent repair to produce four cell lines either homozygous or heterozygous for the PC allele (Table 1). Each of the four lines were cloned by somatic cell nuclear transfer, and full-term pregnancies were established for three of the four lines. In total, five live calves were produced, representing two different dairy genetic backgrounds, 2122 and 2120 (Table 1). When calves were born, a board-certified veterinarian inspected each of the live-born calves visually and by palpation for horn buds, and observed none, which suggested polledness. Analysis of the polled genotype using diagnostic PCR1 confirmed the homozygous introgression of PC into RCI-001, RCI-002 (named Spotigy) and RCI-003 (named Buri), and heterozygous introgression into RCI-004 (RCI, Recombinetics, numbered in order of birth), whereas progenitor cells (2120) were negative for the PC allele (Fig. 1a,b). Two healthy, homozygous polled animals, Spotigy and Buri (Fig. 1c,d and Table 1), which are now more than 10 months old, were phenotypically polled. To evaluate off-target effects in two distinct edited lines, we sequenced the genomes of RCI-001 and RCI-002, derived from clones HP14-B4 and HP7-P4-A1 respectively, along with those of their progenitor cell lines, 2122 and 2120 respectively (Table 1). To avoid dehorning dairy cattle, transcription activator-like effector nucleases (TALENs) were used to edit the genome of bovine embryo fibroblasts with the Pc POLLED allele. After somatic cell nuclear transfer and full-term pregnancies, live calves were screened for the Pc POLLED allele. The results are shown in Figure 1. a. Results: Explain the data in terms of observations and patterns. Be sure to include information about all controls and relate results back to the predicted amplicon sizes. b. Conclusion: Describe how the data support or not support the hypothesis. c. Briefly describe a next experiment to be performed based on the conclusion.
Using figure 1 and the following background information answer the following questions. Identification of the genetic cause of hornlessness in cattle has been the subject of intensive genetic and genomic research, culminating in the nomination of two different candidate neomutations on cattle chromosome 1 that are predicted to have arisen 500-1,000 years ago: a complex allele of Friesian origin (PF), an 80,128 base pair (bp) duplication (1909352–1989480 bp), and a second, simple allele of Celtic origin (PC) corresponding to a duplication of 212 bp (chromosome 1 positions 1705834–1706045) in place of a 10-bp deletion (1706051–1706060)We report the use of genome editing using transcription activator-like effector nucleases (TALENs) to introgress the putative PC POLLED allele into the genome of bovine embryo fibroblasts to try and produce a genotype identical to what is achievable using natural mating, but without the attendant genetic drag and admixture. In our previous studies, we used TALEN-stimulated homology-dependent repair to produce four cell lines either homozygous or heterozygous for the PC allele (Table 1). Each of the four lines were cloned by somatic cell nuclear transfer, and full-term pregnancies were established for three of the four lines. In total, five live calves were produced, representing two different dairy genetic backgrounds, 2122 and 2120 (Table 1). When calves were born, a board-certified veterinarian inspected each of the live-born calves visually and by palpation for horn buds, and observed none, which suggested polledness. Analysis of the polled genotype using diagnostic PCR1 confirmed the homozygous introgression of PC into RCI-001, RCI-002 (named Spotigy) and RCI-003 (named Buri), and heterozygous introgression into RCI-004 (RCI, Recombinetics, numbered in order of birth), whereas progenitor cells (2120) were negative for the PC allele (Fig. 1a,b). Two healthy, homozygous polled animals, Spotigy and Buri (Fig. 1c,d and Table 1), which are now more than 10 months old, were phenotypically polled. To evaluate off-target effects in two distinct edited lines, we sequenced the genomes of RCI-001 and RCI-002, derived from clones HP14-B4 and HP7-P4-A1 respectively, along with those of their progenitor cell lines, 2122 and 2120 respectively (Table 1). To avoid dehorning dairy cattle, transcription activator-like effector nucleases (TALENs) were used to edit the genome of bovine embryo fibroblasts with the Pc POLLED allele. After somatic cell nuclear transfer and full-term pregnancies, live calves were screened for the Pc POLLED allele. The results are shown in Figure 1. a. Results: Explain the data in terms of observations and patterns. Be sure to include information about all controls and relate results back to the predicted amplicon sizes. b. Conclusion: Describe how the data support or not support the hypothesis. c. Briefly describe a next experiment to be performed based on the conclusion.
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ISBN:9781305251052
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Chapter10: From Proteins To Phenotypes
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Using figure 1 and the following background information answer the following questions.
Identification of the genetic cause of hornlessness in cattle has been the subject of intensive genetic and genomic research, culminating in the nomination of two different candidate neomutations on cattle chromosome 1 that are predicted to have arisen 500-1,000 years ago: a complex allele of Friesian origin (PF), an 80,128 base pair (bp) duplication (1909352–1989480 bp), and a second, simple allele of Celtic origin (PC) corresponding to a duplication of 212 bp (chromosome 1 positions 1705834–1706045) in place of a 10-bp deletion (1706051–1706060)We report the use of genome editing using transcription activator-like effector nucleases (TALENs) to introgress the putative PC POLLED allele into the genome of bovine embryo fibroblasts to try and produce a genotype identical to what is achievable using natural mating, but without the attendant genetic drag and admixture. In our previous studies, we used TALEN-stimulated homology-dependent repair to produce four cell lines either homozygous or heterozygous for the PC allele (Table 1). Each of the four lines were cloned by somatic cell nuclear transfer, and full-term pregnancies were established for three of the four lines. In total, five live calves were produced, representing two different dairy genetic backgrounds, 2122 and 2120 (Table 1). When calves were born, a board-certified veterinarian inspected each of the live-born calves visually and by palpation for horn buds, and observed none, which suggested polledness. Analysis of the polled genotype using diagnostic PCR1 confirmed the homozygous introgression of PC into RCI-001, RCI-002 (named Spotigy) and RCI-003 (named Buri), and heterozygous introgression into RCI-004 (RCI, Recombinetics, numbered in order of birth), whereas progenitor cells (2120) were negative for the PC allele (Fig. 1a,b). Two healthy, homozygous polled animals, Spotigy and Buri (Fig. 1c,d and Table 1), which are now more than 10 months old, were phenotypically polled. To evaluate off-target effects in two distinct edited lines, we sequenced the genomes of RCI-001 and RCI-002, derived from clones HP14-B4 and HP7-P4-A1 respectively, along with those of their progenitor cell lines, 2122 and 2120 respectively (Table 1). To avoid dehorning dairy cattle, transcription activator-like effector nucleases (TALENs) were used to edit the genome of bovine embryo fibroblasts with the Pc POLLED allele. After somatic cell nuclear transfer and full-term pregnancies, live calves were screened for the Pc POLLED allele. The results are shown in Figure 1.
a. Results: Explain the data in terms of observations and patterns. Be sure to include information about all controls and relate results back to the predicted amplicon sizes.
b. Conclusion: Describe how the data support or not support the hypothesis.
c. Briefly describe a next experiment to be performed based on the conclusion.
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